13,227 results on '"Caenorhabditis elegans"'
Search Results
2. CRISPR Technology Reveals RAD(51)-ical Mechanisms of Repair in Roundworms: An Educational Primer for Use with "Promotion of Homologous Recombination by SWS-1 in Complex with RAD-51 Paralogs in Caenorhabditis elegans".
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Turcotte CA, Andrews NP, Sloat SA, and Checchi PM
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- Animals, Caenorhabditis elegans Proteins genetics, Rad51 Recombinase genetics, CRISPR-Cas Systems, Caenorhabditis elegans genetics, DNA Repair, Genetics education, Homologous Recombination
- Abstract
The mechanisms cells use to maintain genetic fidelity via DNA repair and the accuracy of these processes have garnered interest from scientists engaged in basic research to clinicians seeking improved treatment for cancer patients. Despite the continued advances, many details of DNA repair are still incompletely understood. In addition, the inherent complexity of DNA repair processes, even at the most fundamental level, makes it a challenging topic. This primer is meant to assist both educators and students in using a recent paper, "Promotion of homologous recombination by SWS-1 in complex with RAD-51 paralogs in Caenorhabditis elegans," to understand mechanisms of DNA repair. The goals of this primer are to highlight and clarify several key techniques utilized, with special emphasis on the clustered, regularly interspaced, short palindromic repeats technique and the ways in which it has revolutionized genetics research, as well as to provide questions for deeper in-class discussion., (Copyright © 2016 by the Genetics Society of America.)
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- 2016
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3. The big in-between. Interview by Kristie Nybo.
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Bargmann C
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- Animals, Caenorhabditis elegans, Humans, Genetics, Neurosciences, Smell
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Cori Bargmann's studies of olfaction and her work on the connections between neural circuits, genes, and behavior caught our attention. Curious to know more, BioTechniques contacted her to find out about the ambition, character, and motivation that led to her success.
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- 2013
4. Predicting phenotypic effects of gene perturbations in C. elegans using an integrated network model.
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Borgwardt K
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- Animals, Caenorhabditis elegans, Computer Simulation, Databases, Factual, Genotype, Humans, Models, Biological, Models, Genetic, Models, Theoretical, Phenotype, RNA Interference, Genetics, Systems Biology
- Abstract
Predicting the phenotype of an organism from its genotype is a central question in genetics. Most importantly, we would like to find out if the perturbation of a single gene may be the cause of a disease. However, our current ability to predict the phenotypic effects of perturbations of individual genes is limited. Network models of genes are one tool for tackling this problem. In a recent study, (Lee et al.) it has been shown that network models covering the majority of genes of an organism can be used for accurately predicting phenotypic effects of gene perturbations in multicellular organisms. ., ((c) 2008 Wiley Periodicals, Inc.)
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- 2008
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5. Roles of Tubulin Concentration during Prometaphase and Ran-GTP during Anaphase of Caenorhabditis elegans Meiosis
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Gong, Ting, McNally, Karen L, Konanoor, Siri, Peraza, Alma, Bailey, Cynthia, Redemann, Stefanie, and McNally, Francis J
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Biochemistry and Cell Biology ,Biological Sciences ,Genetics ,Generic health relevance ,Animals ,Caenorhabditis elegans ,Tubulin ,Spindle Apparatus ,Anaphase ,Caenorhabditis elegans Proteins ,Oocytes ,Prometaphase ,Meiosis ,ran GTP-Binding Protein ,Guanosine Triphosphate ,Chromatin ,Chromosome Segregation ,Biological sciences ,Biomedical and clinical sciences - Abstract
In many animal species, the oocyte meiotic spindle, which is required for chromosome segregation, forms without centrosomes. In some systems, Ran-GEF on chromatin initiates spindle assembly. We found that in Caenorhabditis elegans oocytes, endogenously-tagged Ran-GEF dissociates from chromatin during spindle assembly but re-associates during meiotic anaphase. Meiotic spindle assembly occurred after auxin-induced degradation of Ran-GEF, but anaphase I was faster than controls and extrusion of the first polar body frequently failed. In search of a possible alternative pathway for spindle assembly, we found that soluble tubulin concentrates in the nuclear volume during germinal vesicle breakdown. We found that the concentration of soluble tubulin in the metaphase spindle region is enclosed by ER sheets which exclude cytoplasmic organelles including mitochondria and yolk granules. Measurement of the volume occupied by yolk granules and mitochondria indicated that volume exclusion would be sufficient to explain the concentration of tubulin in the spindle volume. We suggest that this concentration of soluble tubulin may be a redundant mechanism promoting spindle assembly near chromosomes.
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- 2024
6. Clock gene homologs lin-42 and kin-20 regulate circadian rhythms in C. elegans
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Lamberti, Melisa L, Spangler, Rebecca K, Cerdeira, Victoria, Ares, Myriam, Rivollet, Lise, Ashley, Guinevere E, Coronado, Andrea Ramos, Tripathi, Sarvind, Spiousas, Ignacio, Ward, Jordan D, Partch, Carrie L, Bénard, Claire Y, Goya, M Eugenia, and Golombek, Diego A
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Biochemistry and Cell Biology ,Biological Sciences ,Neurosciences ,Sleep Research ,Genetics ,1.1 Normal biological development and functioning ,Underpinning research ,Generic health relevance ,Caenorhabditis elegans ,Animals ,Caenorhabditis elegans Proteins ,Circadian Rhythm ,Mutation ,Circadian Clocks ,Neurons ,CLOCK Proteins ,Gene Expression Regulation ,Transcription Factors - Abstract
Circadian rhythms are endogenous oscillations in nearly all organisms, from prokaryotes to humans, allowing them to adapt to cyclical environments for close to 24 h. Circadian rhythms are regulated by a central clock, based on a transcription-translation feedback loop. One important protein in the central loop in metazoan clocks is PERIOD, which is regulated in part by Casein kinase 1ε/δ (CK1ε/δ) phosphorylation. In the nematode Caenorhabditis elegans, period and casein kinase 1ε/δ are conserved as lin-42 and kin-20, respectively. Here, we studied the involvement of lin-42 and kin-20 in the circadian rhythms of the adult nematode using a bioluminescence-based circadian transcriptional reporter. We show that mutations of lin-42 and kin-20 generate a significantly longer endogenous period, suggesting a role for both genes in the nematode circadian clock, as in other organisms. These phenotypes can be partially rescued by overexpression of either gene under their native promoter. Both proteins are expressed in neurons and epidermal seam cells, as well as in other cells. Depletion of LIN-42 and KIN-20, specifically in neuronal cells after development, was sufficient to lengthen the period of oscillating sur-5 expression. Therefore, we conclude that LIN-42 and KIN-20 are critical regulators of the adult nematode circadian clock through neuronal cells.
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- 2024
7. Screening and characterization of 133 physiologically-relevant environmental chemicals for reproductive toxicity
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Ulaganathan, Gurugowtham, Jiang, Hui, Canio, Noah, Oke, Ashwini, Armstrong, Sujit Silas, Abrahamsson, Dimitri, Varshavsky, Julia R, Lam, Juleen, Cooper, Courtney, Robinson, Joshua F, Fung, Jennifer C, Woodruff, Tracey J, and Allard, Patrick
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Pharmacology and Pharmaceutical Sciences ,Reproductive Medicine ,Biomedical and Clinical Sciences ,Genetics ,Contraception/Reproduction ,Caenorhabditis elegans ,Animals ,Reproduction ,Environmental Pollutants ,Toxicity Tests ,High-Throughput Screening Assays ,C. elegans ,Reproductive toxicity ,NAMs ,Alternative testing ,QACs ,Paediatrics and Reproductive Medicine ,Public Health and Health Services ,Toxicology ,Pharmacology and pharmaceutical sciences ,Reproductive medicine - Abstract
Reproduction is a functional outcome that relies on complex cellular, tissue, and organ interactions that span the developmental period to adulthood. Thus, the assessment of its disruption by environmental chemicals would benefit significantly from scalable and innovative approaches to testing using functionally comparable reproductive models such as the nematode C. elegans. We adapted a previously described low-throughput in vivo chromosome segregation assay using C. elegans predictive of reproductive toxicity and leveraged available public data sources (ToxCast, ICE) to screen and characterize 133 physiologically-relevant chemicals in a high-throughput manner. The screening outcome was further validated in a second, independent in vivo assay assessing embryonic viability. In total, 13 chemicals were classified as reproductive toxicants with the two most active chemicals belonging to the large family of Quaternary Ammonium Compounds (QACs) commonly used as disinfectants but with limited available reproductive toxicity data. We compared the results from the C. elegans assay with ToxCast in vitro data compiled from 700+ cell response assays and 300+ signaling pathways-based assays. We did not observe a difference in the bioactivity or in the average potency (AC50) between the top and bottom chemicals. However, the intended target categories were significantly different between the classified chemicals with, in particular, an over-representation of steroid hormone targets for the high Z-score chemicals. Taken together, these results point to the value of in vivo models that scale to high-throughput level for reproductive toxicity assessment and to the need to prioritize the assessment of QACs impacts on reproduction.
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- 2024
8. Katanin, kinesin-13, and ataxin-2 inhibit premature interaction between maternal and paternal genomes in C. elegans zygotes
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Beath, Elizabeth A, Bailey, Cynthia, Magadam, Meghana Mahantesh, Qiu, Shuyan, McNally, Karen L, and McNally, Francis J
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Reproductive Medicine ,Biomedical and Clinical Sciences ,Biological Sciences ,Contraception/Reproduction ,Genetics ,Reproductive health and childbirth ,Animals ,Caenorhabditis elegans ,Katanin ,Zygote ,Kinesins ,Male ,Ataxin-2 ,Caenorhabditis elegans Proteins ,Spermatozoa ,Female ,Fertilization ,meiosis ,fertilization ,sperm ,C. elegans ,cell biology ,Biochemistry and Cell Biology ,Biological sciences ,Biomedical and clinical sciences ,Health sciences - Abstract
Fertilization occurs before the completion of oocyte meiosis in the majority of animal species and sperm contents move long distances within the zygotes of mouse and C. elegans. If incorporated into the meiotic spindle, paternal chromosomes could be expelled into a polar body resulting in lethal monosomy. Through live imaging of fertilization in C. elegans, we found that the microtubule disassembling enzymes, katanin and kinesin-13 limit long-range movement of sperm contents and that maternal ataxin-2 maintains paternal DNA and paternal mitochondria as a cohesive unit that moves together. Depletion of katanin or double depletion of kinesin-13 and ataxin-2 resulted in the capture of the sperm contents by the meiotic spindle. Thus limiting movement of sperm contents and maintaining cohesion of sperm contents within the zygote both contribute to preventing premature interaction between maternal and paternal genomes.
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- 2024
9. Proteasome inhibition triggers tissue-specific immune responses against different pathogens in C. elegans
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Grover, Manish, Gang, Spencer S, Troemel, Emily R, and Barkoulas, Michalis
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Medical Microbiology ,Biomedical and Clinical Sciences ,Biological Sciences ,Infectious Diseases ,Prevention ,Emerging Infectious Diseases ,Vaccine Related ,Genetics ,Aetiology ,2.1 Biological and endogenous factors ,Infection ,Animals ,Caenorhabditis elegans ,Transcription Factors ,DNA-Binding Proteins ,Proteasome Endopeptidase Complex ,Caenorhabditis elegans Proteins ,Immunity ,Innate ,Bacterial Infections ,Agricultural and Veterinary Sciences ,Medical and Health Sciences ,Developmental Biology ,Agricultural ,veterinary and food sciences ,Biological sciences ,Biomedical and clinical sciences - Abstract
Protein quality control pathways play important roles in resistance against pathogen infection. For example, the conserved transcription factor SKN-1/NRF up-regulates proteostasis capacity after blockade of the proteasome and also promotes resistance against bacterial infection in the nematode Caenorhabditis elegans. SKN-1/NRF has 3 isoforms, and the SKN-1A/NRF1 isoform, in particular, regulates proteasomal gene expression upon proteasome dysfunction as part of a conserved bounce-back response. We report here that, in contrast to the previously reported role of SKN-1 in promoting resistance against bacterial infection, loss-of-function mutants in skn-1a and its activating enzymes ddi-1 and png-1 show constitutive expression of immune response programs against natural eukaryotic pathogens of C. elegans. These programs are the oomycete recognition response (ORR), which promotes resistance against oomycetes that infect through the epidermis, and the intracellular pathogen response (IPR), which promotes resistance against intestine-infecting microsporidia. Consequently, skn-1a mutants show increased resistance to both oomycete and microsporidia infections. We also report that almost all ORR/IPR genes induced in common between these programs are regulated by the proteasome and interestingly, specific ORR/IPR genes can be induced in distinct tissues depending on the exact trigger. Furthermore, we show that increasing proteasome function significantly reduces oomycete-mediated induction of multiple ORR markers. Altogether, our findings demonstrate that proteasome regulation keeps innate immune responses in check in a tissue-specific manner against natural eukaryotic pathogens of the C. elegans epidermis and intestine.
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- 2024
10. Characterizing human KIF1Bβ motor activity by single-molecule motility assays and Caenorhabditis elegans genetics.
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Rei Iguchi, Tomoki Kita, Taisei Watanabe, Kyoko Chiba, and Shinsuke Niwa
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AXONAL transport , *SYNAPTIC vesicles , *CAENORHABDITIS elegans , *NEUROLOGICAL disorders , *GENETICS - Abstract
The axonal transport of synaptic vesicle precursors relies on KIF1A and UNC-104 ortholog motors. In mammals, KIF1Bβ is also responsible for the axonal transport of synaptic vesicle precursors. Mutations in KIF1A and KIF1Bβ lead to a wide range of neuropathies. Although previous studies have revealed the biochemical, biophysical and cell biological properties of KIF1A, and its defects in neurological disorders, the fundamental properties of KIF1Bβ remain elusive. In this study, we determined the motile parameters of KIF1Bβ through single-molecule motility assays. We found that the C-terminal region of KIF1Bβ has an inhibitory role in motor activity. AlphaFold2 prediction suggests that the C-terminal region blocks the motor domain. Additionally, we established simple methods for testing the axonal transport activity of human KIF1Bβ using Caenorhabditis elegans genetics. Taking advantage of these methods, we demonstrated that these assays enable the detection of reduced KIF1Bβ activities, both in vitro and in vivo, caused by a Charcot–Marie–Tooth disease-associated Q98L mutation. [ABSTRACT FROM AUTHOR]
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- 2024
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11. Overexpression of mitochondrial fission or mitochondrial fusion genes enhances resilience and extends longevity.
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Traa, Annika, Keil, Allison, AlOkda, Abdelrahman, Jacob‐Tomas, Suleima, Tamez González, Aura A., Zhu, Shusen, Rudich, Zenith, and Van Raamsdonk, Jeremy M.
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MITOCHONDRIAL dynamics , *GENE fusion , *GENETICS , *CAENORHABDITIS elegans , *MORPHOLOGY - Abstract
The dynamicity of the mitochondrial network is crucial for meeting the ever‐changing metabolic and energy needs of the cell. Mitochondrial fission promotes the degradation and distribution of mitochondria, while mitochondrial fusion maintains mitochondrial function through the complementation of mitochondrial components. Previously, we have reported that mitochondrial networks are tubular, interconnected, and well‐organized in young, healthy C. elegans, but become fragmented and disorganized with advancing age and in models of age‐associated neurodegenerative disease. In this work, we examine the effects of increasing mitochondrial fission or mitochondrial fusion capacity by ubiquitously overexpressing the mitochondrial fission gene drp‐1 or the mitochondrial fusion genes fzo‐1 and eat‐3, individually or in combination. We then measured mitochondrial function, mitochondrial network morphology, physiologic rates, stress resistance, and lifespan. Surprisingly, we found that overexpression of either mitochondrial fission or fusion machinery both resulted in an increase in mitochondrial fragmentation. Similarly, both mitochondrial fission and mitochondrial fusion overexpression strains have extended lifespans and increased stress resistance, which in the case of the mitochondrial fusion overexpression strains appears to be at least partially due to the upregulation of multiple pathways of cellular resilience in these strains. Overall, our work demonstrates that increasing the expression of mitochondrial fission or fusion genes extends lifespan and improves biological resilience without promoting the maintenance of a youthful mitochondrial network morphology. This work highlights the importance of the mitochondria for both resilience and longevity. [ABSTRACT FROM AUTHOR]
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- 2024
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12. Direct inference of the distribution of fitness effects of spontaneous mutations from recombinant inbred Caenorhabditis elegans mutation accumulation lines.
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Crombie, Timothy A, Rajaei, Moein, Saxena, Ayush Shekhar, Johnson, Lindsay M, Saber, Sayran, Tanny, Robyn E, Ponciano, José Miguel, Andersen, Erik C, Zhou, Juannan, and Baer, Charles F
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BIOLOGICAL models , *RESEARCH funding , *CONSANGUINITY , *ANIMAL experimentation , *GENETIC mutation , *CAENORHABDITIS elegans , *GENETIC techniques , *GENETICS , *HAPLOTYPES - Abstract
The distribution of fitness effects of new mutations plays a central role in evolutionary biology. Estimates of the distribution of fitness effect from experimental mutation accumulation lines are compromised by the complete linkage disequilibrium between mutations in different lines. To reduce the linkage disequilibrium, we constructed 2 sets of recombinant inbred lines from a cross of 2 Caenorhabditis elegans mutation accumulation lines. One set of lines ("RIAILs") was intercrossed for 10 generations prior to 10 generations of selfing; the second set of lines ("RILs") omitted the intercrossing. Residual linkage disequilibrium in the RIAILs is much less than in the RILs, which affects the inferred distribution of fitness effect when the sets of lines are analyzed separately. The best-fit model estimated from all lines (RIAILs + RILs) infers a large fraction of mutations with positive effects (∼40%); models that constrain mutations to have negative effects fit much worse. The conclusion is the same using only the RILs. For the RIAILs, however, models that constrain mutations to have negative effects fit nearly as well as models that allow positive effects. When mutations in high linkage disequilibrium are pooled into haplotypes, the inferred distribution of fitness effect becomes increasingly negative-skewed and leptokurtic. We conclude that the conventional wisdom—most mutations have effects near 0, a handful of mutations have effects that are substantially negative, and mutations with positive effects are very rare—is likely correct, and that unless it can be shown otherwise, estimates of the distribution of fitness effect that infer a substantial fraction of mutations with positive effects are likely confounded by linkage disequilibrium. [ABSTRACT FROM AUTHOR]
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- 2024
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13. Neurogenesis in Caenorhabditis elegans.
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Poole, Richard J, Flames, Nuria, and Cochella, Luisa
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NEURAL physiology , *NEURAL development , *NEURONS , *NEUROPLASTICITY , *NEUROPHYSIOLOGY , *TRANSCRIPTION factors , *GENES , *GENE expression , *CHROMOSOMES , *CAENORHABDITIS elegans , *STEM cells , *GENETICS - Abstract
Animals rely on their nervous systems to process sensory inputs, integrate these with internal signals, and produce behavioral outputs. This is enabled by the highly specialized morphologies and functions of neurons. Neuronal cells share multiple structural and physiological features, but they also come in a large diversity of types or classes that give the nervous system its broad range of functions and plasticity. This diversity, first recognized over a century ago, spurred classification efforts based on morphology, function, and molecular criteria. Caenorhabditis elegans , with its precisely mapped nervous system at the anatomical level, an extensive molecular description of most of its neurons, and its genetic amenability, has been a prime model for understanding how neurons develop and diversify at a mechanistic level. Here, we review the gene regulatory mechanisms driving neurogenesis and the diversification of neuron classes and subclasses in C. elegans. We discuss our current understanding of the specification of neuronal progenitors and their differentiation in terms of the transcription factors involved and ensuing changes in gene expression and chromatin landscape. The central theme that has emerged is that the identity of a neuron is defined by modules of gene batteries that are under control of parallel yet interconnected regulatory mechanisms. We focus on how, to achieve these terminal identities, cells integrate information along their developmental lineages. Moreover, we discuss how neurons are diversified postembryonically in a time-, genetic sex-, and activity-dependent manner. Finally, we discuss how the understanding of neuronal development can provide insights into the evolution of neuronal diversity. [ABSTRACT FROM AUTHOR]
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- 2024
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14. Conditional nmy-1 and nmy-2 alleles establish that nonmuscle myosins are required for late Caenorhabditis elegans embryonic elongation.
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Molnar, Kelly, Suman, Shashi Kumar, Eichelbrenner, Jeanne, Plancke, Camille N, Robin, François B, and Labouesse, Michel
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MUSCLE physiology , *PROTEIN metabolism , *EMBRYOS , *MEDICAL protocols , *MUSCLE proteins , *MYOSIN , *EPIDERMIS , *RNA , *CAENORHABDITIS elegans , *TEMPERATURE , *GENETIC mutation , *MICROSCOPY , *ALLELES , *GENETICS , *GENOTYPES - Abstract
The elongation of Caenorhabditis elegans embryos allows examination of mechanical interactions between adjacent tissues. Muscle contractions during late elongation induce the remodeling of epidermal circumferential actin filaments through mechanotransduction. Force inputs from the muscles deform circumferential epidermal actin filament, which causes them to be severed, eventually reformed, and shortened. This squeezing force drives embryonic elongation. We investigated the possible role of the nonmuscle myosins NMY-1 and NMY-2 in this process using nmy-1 and nmy-2 thermosensitive alleles. Our findings show these myosins act redundantly in late elongation, since double nmy-2(ts); nmy-1(ts) mutants immediately stop elongation when raised to 25°C. Their inactivation does not reduce muscle activity, as measured from epidermis deformation, suggesting that they are directly involved in the multistep process of epidermal remodeling. Furthermore, NMY-1 and NMY-2 inactivation is reversible when embryos are kept at the nonpermissive temperature for a few hours. However, after longer exposure to 25°C double mutant embryos fail to resume elongation, presumably because NMY-1 was seen to form protein aggregates. We propose that the two C. elegans nonmuscle myosin II act during actin remodeling either to bring severed ends or hold them. [ABSTRACT FROM AUTHOR]
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- 2024
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15. The chromatin-associated 53BP1 ortholog, HSR-9, regulates recombinational repair and X chromosome segregation in the Caenorhabditis elegans germ line.
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Li, Qianyan, Hariri, Sara, Calidas, Aashna, Kaur, Arshdeep, Huey, Erica, and Engebrecht, JoAnne
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BIOLOGICAL models , *GERM cells , *RESEARCH funding , *CELL physiology , *CELL proliferation , *DESCRIPTIVE statistics , *MICE , *CHROMOSOMES , *DNA damage , *DNA repair , *ANIMAL experimentation , *CAENORHABDITIS elegans , *GENETIC mutation , *DNA-binding proteins , *GENETICS - Abstract
53BP1 plays a crucial role in regulating DNA damage repair pathway choice and checkpoint signaling in somatic cells; however, its role in meiosis has remained enigmatic. In this study, we demonstrate that the Caenorhabditis elegans ortholog of 53BP1, HSR-9 , associates with chromatin in both proliferating and meiotic germ cells. Notably, HSR-9 is enriched on the X chromosome pair in pachytene oogenic germ cells. HSR-9 is also present at kinetochores during both mitotic and meiotic divisions but does not appear to be essential for monitoring microtubule–kinetochore attachments or tension. Using cytological markers of different steps in recombinational repair, we found that HSR-9 influences the processing of a subset of meiotic double-stranded breaks into COSA-1 -marked crossovers. Additionally, HSR-9 plays a role in meiotic X chromosome segregation under conditions where X chromosomes fail to pair, synapse, and recombine. Together, these results highlight that chromatin-associated HSR-9 has both conserved and unique functions in the regulation of meiotic chromosome behavior. [ABSTRACT FROM AUTHOR]
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- 2024
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16. Protocol for nuclear dissociation of the adult C. elegans for single-nucleus RNA sequencing and its application for mapping environmental responses
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Levenson, Max T, Barrere-Cain, Rio, Truong, Lisa, Chen, Yen-Wei, Shuck, Karissa, Panter, Blake, Reich, Ella, Yang, Xia, and Allard, Patrick
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Biological Sciences ,Genetics ,Human Genome ,1.1 Normal biological development and functioning ,Generic health relevance ,Animals ,Caenorhabditis elegans ,Sequence Analysis ,RNA ,Base Sequence ,Cell Nucleus ,RNA ,Small Nuclear ,Gene Expression ,Model Organisms ,RNA-seq ,Single Cell - Abstract
Caenorhabditis elegans is a valuable model to study organ, tissue, and cell-type responses to external cues. However, the nematode comprises multiple syncytial tissues with spatial coordinates corresponding to distinct nuclear transcriptomes. Here, we present a single-nucleus RNA sequencing (snRNA-seq) protocol that aims to overcome difficulties encountered with single-cell RNA sequencing in C. elegans. We describe steps for isolating C. elegans nuclei for downstream applications including snRNA-seq applied to the context of alcohol exposure. For complete details on the use and execution of this protocol, please refer to Truong et al. (2023).1.
- Published
- 2023
17. Key processes required for the different stages of fungal carnivory by a nematode-trapping fungus
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Lin, Hung-Che, de Ulzurrun, Guillermo Vidal-Diez, Chen, Sheng-An, Yang, Ching-Ting, Tay, Rebecca J, Iizuka, Tomoyo, Huang, Tsung-Yu, Kuo, Chih-Yen, Gonçalves, A Pedro, Lin, Siou-Ying, Chang, Yu-Chu, Stajich, Jason E, Schwarz, Erich M, and Hsueh, Yen-Ping
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Microbiology ,Biological Sciences ,Genetics ,Biotechnology ,Underpinning research ,1.1 Normal biological development and functioning ,Generic health relevance ,Animals ,Caenorhabditis elegans ,Carnivory ,Nematoda ,Gene Expression Profiling ,Metalloproteases ,Agricultural and Veterinary Sciences ,Medical and Health Sciences ,Developmental Biology ,Agricultural ,veterinary and food sciences ,Biological sciences ,Biomedical and clinical sciences - Abstract
Nutritional deprivation triggers a switch from a saprotrophic to predatory lifestyle in soil-dwelling nematode-trapping fungi (NTF). In particular, the NTF Arthrobotrys oligospora secretes food and sex cues to lure nematodes to its mycelium and is triggered to develop specialized trapping devices. Captured nematodes are then invaded and digested by the fungus, thus serving as a food source. In this study, we examined the transcriptomic response of A. oligospora across the stages of sensing, trap development, and digestion upon exposure to the model nematode Caenorhabditis elegans. A. oligospora enacts a dynamic transcriptomic response, especially of protein secretion-related genes, in the presence of prey. Two-thirds of the predicted secretome of A. oligospora was up-regulated in the presence of C. elegans at all time points examined, and among these secreted proteins, 38.5% are predicted to be effector proteins. Furthermore, functional studies disrupting the t-SNARE protein Sso2 resulted in impaired ability to capture nematodes. Additionally, genes of the DUF3129 family, which are expanded in the genomes of several NTF, were highly up-regulated upon nematode exposure. We observed the accumulation of highly expressed DUF3129 proteins in trap cells, leading us to name members of this gene family as Trap Enriched Proteins (TEPs). Gene deletion of the most highly expressed TEP gene, TEP1, impairs the function of traps and prevents the fungus from capturing prey efficiently. In late stages of predation, we observed up-regulation of a variety of proteases, including metalloproteases. Following penetration of nematodes, these metalloproteases facilitate hyphal growth required for colonization of prey. These findings provide insights into the biology of the predatory lifestyle switch in a carnivorous fungus and provide frameworks for other fungal-nematode predator-prey systems.
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- 2023
18. Regulation of defective mitochondrial DNA accumulation and transmission in C. elegans by the programmed cell death and aging pathways.
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Flowers, Sagen, Kothari, Rushali, Torres Cleuren, Yamila, Alcorn, Melissa, Ewe, Chee, Alok, Geneva, Fiallo, Samantha, Joshi, Pradeep, and Rothman, Joel
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C. elegans ,aging ,cell biology ,genetics ,genomics ,heteroplasmy ,insulin signaling ,programmed cell death ,purifying selection ,uaDf5 ,Animals ,Caenorhabditis elegans ,DNA ,Mitochondrial ,Apoptosis ,Caspases ,Caenorhabditis elegans Proteins ,Aging - Abstract
The heteroplasmic state of eukaryotic cells allows for cryptic accumulation of defective mitochondrial genomes (mtDNA). Purifying selection mechanisms operate to remove such dysfunctional mtDNAs. We found that activators of programmed cell death (PCD), including the CED-3 and CSP-1 caspases, the BH3-only protein CED-13, and PCD corpse engulfment factors, are required in C. elegans to attenuate germline abundance of a 3.1-kb mtDNA deletion mutation, uaDf5, which is normally stably maintained in heteroplasmy with wildtype mtDNA. In contrast, removal of CED-4/Apaf1 or a mutation in the CED-4-interacting prodomain of CED-3, do not increase accumulation of the defective mtDNA, suggesting induction of a non-canonical germline PCD mechanism or non-apoptotic action of the CED-13/caspase axis. We also found that the abundance of germline mtDNAuaDf5 reproducibly increases with age of the mothers. This effect is transmitted to the offspring of mothers, with only partial intergenerational removal of the defective mtDNA. In mutants with elevated mtDNAuaDf5 levels, this removal is enhanced in older mothers, suggesting an age-dependent mechanism of mtDNA quality control. Indeed, we found that both steady-state and age-dependent accumulation rates of uaDf5 are markedly decreased in long-lived, and increased in short-lived, mutants. These findings reveal that regulators of both PCD and the aging program are required for germline mtDNA quality control and its intergenerational transmission.
- Published
- 2023
19. Methylglyoxal-derived hydroimidazolone, MG-H1, increases food intake by altering tyramine signaling via the GATA transcription factor ELT-3 in Caenorhabditis elegans.
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Muthaiyan Shanmugam, Muniesh, Chaudhuri, Jyotiska, Sellegounder, Durai, Sahu, Amit, Guha, Sanjib, Chamoli, Manish, Hodge, Brian, Bose, Neelanjan, Roberts, Charis, Farrera, Dominique, Lithgow, Gordon, Galligan, James, Kapahi, Pankaj, and Sarpong, Richmond
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C. elegans ,advanced glycation end-products ,elt-3 ,feeding ,genetics ,genomics ,glod-4 ,neuroscience ,pharyngeal pumping ,tyramine ,Animals ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Pyruvaldehyde ,Magnesium Oxide ,GATA Transcription Factors ,Signal Transduction ,Tyramine ,Glycation End Products ,Advanced ,Eating - Abstract
The Maillard reaction, a chemical reaction between amino acids and sugars, is exploited to produce flavorful food ubiquitously, from the baking industry to our everyday lives. However, the Maillard reaction also occurs in all cells, from prokaryotes to eukaryotes, forming advanced glycation end-products (AGEs). AGEs are a heterogeneous group of compounds resulting from the irreversible reaction between biomolecules and α-dicarbonyls (α-DCs), including methylglyoxal (MGO), an unavoidable byproduct of anaerobic glycolysis and lipid peroxidation. We previously demonstrated that Caenorhabditis elegans mutants lacking the glod-4 glyoxalase enzyme displayed enhanced accumulation of α-DCs, reduced lifespan, increased neuronal damage, and touch hypersensitivity. Here, we demonstrate that glod-4 mutation increased food intake and identify that MGO-derived hydroimidazolone, MG-H1, is a mediator of the observed increase in food intake. RNAseq analysis in glod-4 knockdown worms identified upregulation of several neurotransmitters and feeding genes. Suppressor screening of the overfeeding phenotype identified the tdc-1-tyramine-tyra-2/ser-2 signaling as an essential pathway mediating AGE (MG-H1)-induced feeding in glod-4 mutants. We also identified the elt-3 GATA transcription factor as an essential upstream regulator for increased feeding upon accumulation of AGEs by partially controlling the expression of tdc-1 gene. Furthermore, the lack of either tdc-1 or tyra-2/ser-2 receptors suppresses the reduced lifespan and rescues neuronal damage observed in glod-4 mutants. Thus, in C. elegans, we identified an elt-3 regulated tyramine-dependent pathway mediating the toxic effects of MG-H1 AGE. Understanding this signaling pathway may help understand hedonistic overfeeding behavior observed due to modern AGE-rich diets.
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- 2023
20. Gene duplication and evolutionary plasticity of lin-12/Notch gene function in Caenorhabditis.
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Lyu, Haimeng, Moya, Nicolas D, Andersen, Erik C, and Chamberlin, Helen M
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RESEARCH funding , *CELLULAR signal transduction , *MAXIMUM likelihood statistics , *DESCRIPTIVE statistics , *GENE expression , *GENOME editing , *GENETIC mutation , *CAENORHABDITIS elegans , *MICROSCOPY , *SEQUENCE analysis , *GENOMES , *GENETICS , *ALLELES , *PHENOTYPES - Abstract
Gene duplication is an important substrate for the evolution of new gene functions, but the impacts of gene duplicates on their own activities and on the developmental networks in which they act are poorly understood. Here, we use a natural experiment of lin-12 /Notch gene duplication within the nematode genus Caenorhabditis , combined with characterization of loss- and gain-of-function mutations, to uncover functional distinctions between the duplicate genes in 1 species (Caenorhabditis briggsae) and their single-copy ortholog in Caenorhabditis elegans. First, using improved genomic sequence and gene model characterization, we confirm that the C. briggsae genome includes 2 complete lin-12 genes, whereas most other genes encoding proteins that participate in the LIN-12 signaling pathway retain a one-to-one orthology with C. elegans. We use CRISPR-mediated genome editing to introduce alleles predicted to cause gain-of-function (gf) or loss-of-function (lf) into each C. briggsae gene and find that the gf mutations uncover functional distinctions not apparent from the lf alleles. Specifically, Cbr-lin-12.1 (gf) , but not Cbr-lin-12.2 (gf) , causes developmental defects similar to those observed in Cel-lin-12 (gf). In contrast to Cel-lin-12 (gf) , however, the Cbr-lin-12.1 (gf) alleles do not cause dominant phenotypes as compared to the wild type, and the mutant phenotype is observed only when 2 gf alleles are present. Our results demonstrate that gene duplicates can exhibit differential capacities to compensate for each other and to interfere with normal development, and uncover coincident gene duplication and evolution of developmental sensitivity to LIN-12 /Notch activity. [ABSTRACT FROM AUTHOR]
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- 2024
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21. Dendrite morphogenesis in Caenorhabditis elegans.
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Heiman, Maxwell G and Bülow, Hannes E
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PROTEIN metabolism , *SOMATOSENSORY disorders , *TISSUES , *MORPHOGENESIS , *NEUROGLIA , *NEURONS , *TISSUE adhesions , *BIOCHEMISTRY , *CAENORHABDITIS elegans , *SENSORY receptors , *GENETICS - Abstract
Since the days of Ramón y Cajal, the vast diversity of neuronal and particularly dendrite morphology has been used to catalog neurons into different classes. Dendrite morphology varies greatly and reflects the different functions performed by different types of neurons. Significant progress has been made in our understanding of how dendrites form and the molecular factors and forces that shape these often elaborately sculpted structures. Here, we review work in the nematode Caenorhabditis elegans that has shed light on the developmental mechanisms that mediate dendrite morphogenesis with a focus on studies investigating ciliated sensory neurons and the highly elaborated dendritic trees of somatosensory neurons. These studies, which combine time-lapse imaging, genetics, and biochemistry, reveal an intricate network of factors that function both intrinsically in dendrites and extrinsically from surrounding tissues. Therefore, dendrite morphogenesis is the result of multiple tissue interactions, which ultimately determine the shape of dendritic arbors. [ABSTRACT FROM AUTHOR]
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- 2024
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- View/download PDF
22. Understanding the factors regulating host–microbiome interactions using Caenorhabditis elegans.
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Singh, Anupama and Luallen, Robert J.
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CAENORHABDITIS elegans , *GENETIC engineering , *GUT microbiome , *COLONIZATION (Ecology) , *HUMAN microbiota , *GENETIC variation - Abstract
The Human Microbiome Project was a research programme that successfully identified associations between microbial species and healthy or diseased individuals. However, a major challenge identified was the absence of model systems for studying host–microbiome interactions, which would increase our capacity to uncover molecular interactions, understand organ-specificity and discover new microbiome-altering health interventions. Caenorhabditis elegans has been a pioneering model organism for over 70 years but was largely studied in the absence of a microbiome. Recently, ecological sampling of wild nematodes has uncovered a large amount of natural genetic diversity as well as a slew of associated microbiota. The field has now explored the interactions of C. elegans with its associated gut microbiome, a defined and non-random microbial community, highlighting its suitability for dissecting host–microbiome interactions. This core microbiome is being used to study the impact of host genetics, age and stressors on microbiome composition. Furthermore, single microbiome species are being used to dissect molecular interactions between microbes and the animal gut. Being amenable to health altering genetic and non-genetic interventions, C. elegans has emerged as a promising system to generate and test new hypotheses regarding host–microbiome interactions, with the potential to uncover novel paradigms relevant to other systems. This article is part of the theme issue 'Sculpting the microbiome: how host factors determine and respond to microbial colonization'. [ABSTRACT FROM AUTHOR]
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- 2024
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23. WormBase 2024: status and transitioning to Alliance infrastructure.
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Sternberg, Paul W, Auken, Kimberly Van, Wang, Qinghua, Wright, Adam, Yook, Karen, Zarowiecki, Magdalena, Arnaboldi, Valerio, Becerra, Andrés, Brown, Stephanie, Cain, Scott, Chan, Juancarlos, Chen, Wen J, Cho, Jaehyoung, Davis, Paul, Diamantakis, Stavros, Dyer, Sarah, Grigoriadis, Dionysis, Grove, Christian A, Harris, Todd, and Howe, Kevin
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DATABASES , *GENOMICS , *INFORMATION resources , *BIOINFORMATICS , *CAENORHABDITIS elegans , *GENETICS - Abstract
WormBase has been the major repository and knowledgebase of information about the genome and genetics of Caenorhabditis elegans and other nematodes of experimental interest for over 2 decades. We have 3 goals: to keep current with the fast-paced C. elegans research, to provide better integration with other resources, and to be sustainable. Here, we discuss the current state of WormBase as well as progress and plans for moving core WormBase infrastructure to the Alliance of Genome Resources (the Alliance). As an Alliance member, WormBase will continue to interact with the C. elegans community, develop new features as needed, and curate key information from the literature and large-scale projects. [ABSTRACT FROM AUTHOR]
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- 2024
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24. Updates to the Alliance of Genome Resources central infrastructure.
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Consortium, The Alliance of Genome Resources
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BIOLOGICAL models , *DATABASES , *COMPUTER software , *DATA mining , *DATABASE management , *DATA curation , *ARTIFICIAL intelligence , *INFORMATION resources , *FISHES , *PROFESSIONS , *MICE , *RATS , *INFORMATION services , *INFORMATION retrieval , *CAENORHABDITIS elegans , *INSECTS , *ONTOLOGIES (Information retrieval) , *MACHINE learning , *GENOMES , *GENETICS , *YEAST , *ANURA - Abstract
The Alliance of Genome Resources (Alliance) is an extensible coalition of knowledgebases focused on the genetics and genomics of intensively studied model organisms. The Alliance is organized as individual knowledge centers with strong connections to their research communities and a centralized software infrastructure, discussed here. Model organisms currently represented in the Alliance are budding yeast, Caenorhabditis elegans , Drosophila , zebrafish, frog, laboratory mouse, laboratory rat, and the Gene Ontology Consortium. The project is in a rapid development phase to harmonize knowledge, store it, analyze it, and present it to the community through a web portal, direct downloads, and application programming interfaces (APIs). Here, we focus on developments over the last 2 years. Specifically, we added and enhanced tools for browsing the genome (JBrowse), downloading sequences, mining complex data (AllianceMine), visualizing pathways, full-text searching of the literature (Textpresso), and sequence similarity searching (SequenceServer). We enhanced existing interactive data tables and added an interactive table of paralogs to complement our representation of orthology. To support individual model organism communities, we implemented species-specific "landing pages" and will add disease-specific portals soon; in addition, we support a common community forum implemented in Discourse software. We describe our progress toward a central persistent database to support curation, the data modeling that underpins harmonization, and progress toward a state-of-the-art literature curation system with integrated artificial intelligence and machine learning (AI/ML). [ABSTRACT FROM AUTHOR]
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- 2024
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25. Whole-body gene expression atlas of an adult metazoan
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Ghaddar, Abbas, Armingol, Erick, Huynh, Chau, Gevirtzman, Louis, Lewis, Nathan E, Waterston, Robert, and O’Rourke, Eyleen J
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Genetics ,1.1 Normal biological development and functioning ,Underpinning research ,Generic health relevance ,Animals ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Transcription Factors ,Gene Expression Regulation ,Gene Expression - Abstract
Gene activity defines cell identity, drives intercellular communication, and underlies the functioning of multicellular organisms. We present the single-cell resolution atlas of gene activity of a fertile adult metazoan: Caenorhabditis elegans. This compendium comprises 180 distinct cell types and 19,657 expressed genes. We predict 7541 transcription factor expression profile associations likely responsible for defining cellular identity. We predict thousands of intercellular interactions across the C. elegans body and the ligand-receptor pairs that mediate them, some of which we experimentally validate. We identify 172 genes that show consistent expression across cell types, are involved in basic and essential functions, and are conserved across phyla; therefore, we present them as experimentally validated housekeeping genes. We developed the WormSeq application to explore these data. In addition to the integrated gene-to-systems biology, we present genome-scale single-cell resolution testable hypotheses that we anticipate will advance our understanding of the molecular mechanisms, underlying the functioning of a multicellular organism and the perturbations that lead to its malfunction.
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- 2023
26. Single-nucleus resolution mapping of the adult C. elegans and its application to elucidate inter- and trans-generational response to alcohol
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Truong, Lisa, Chen, Yen-Wei, Barrere-Cain, Rio, Levenson, Max T, Shuck, Karissa, Xiao, Wen, da Veiga Beltrame, Eduardo, Panter, Blake, Reich, Ella, Sternberg, Paul W, Yang, Xia, and Allard, Patrick
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Biological Sciences ,Genetics ,Substance Misuse ,Human Genome ,Alcoholism ,Alcohol Use and Health ,1.1 Normal biological development and functioning ,2.1 Biological and endogenous factors ,Generic health relevance ,Good Health and Well Being ,Animals ,Adult ,Humans ,Caenorhabditis elegans ,Transcriptome ,Gene Expression Profiling ,Ethanol ,RNA ,Small Nuclear ,C. elegans ,CP: Molecular biology ,alcohol ,ethanol ,germline ,snRNA-seq ,Biochemistry and Cell Biology ,Medical Physiology ,Biological sciences - Abstract
Single-cell transcriptomic platforms provide an opportunity to map an organism's response to environmental cues with high resolution. Here, we applied single-nucleus RNA sequencing (snRNA-seq) to establish the tissue and cell type-resolved transcriptome of the adult C. elegans and characterize the inter- and trans-generational transcriptional impact of ethanol. We profiled the transcriptome of 41,749 nuclei resolving into 31 clusters, representing a diverse array of adult cell types including syncytial tissues. Following exposure to human-relevant doses of alcohol, several germline, striated muscle, and neuronal clusters were identified as being the most transcriptionally impacted at the F1 and F3 generations. The effect on germline clusters was confirmed by phenotypic enrichment analysis as well as by functional validation, which revealed a remarkable inter- and trans-generational increase in germline apoptosis, aneuploidy, and embryonic lethality. Together, snRNA-seq represents a valuable approach for the detailed examination of an adult organism's response to environmental exposures.
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- 2023
27. NHR-23 activity is necessary for C. elegans developmental progression and apical extracellular matrix structure and function
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Johnson, Londen C, Vo, An A, Clancy, John C, Myles, Krista M, Pooranachithra, Murugesan, Aguilera, Joseph, Levenson, Max T, Wohlenberg, Chloe, Rechtsteiner, Andreas, Ragle, James Matthew, Chisholm, Andrew D, and Ward, Jordan D
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Biochemistry and Cell Biology ,Biomedical and Clinical Sciences ,Biological Sciences ,Genetics ,Animals ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Epithelium ,Extracellular Matrix ,Receptors ,Cytoplasmic and Nuclear ,C. elegans ,Molting ,NHR-23 ,Nuclear hormone receptor ,Apical extracellular matrix ,Auxin-inducible degron ,C. elegans ,Medical and Health Sciences ,Biological sciences ,Biomedical and clinical sciences ,Health sciences - Abstract
Nematode molting is a remarkable process where animals must repeatedly build a new apical extracellular matrix (aECM) beneath a previously built aECM that is subsequently shed. The nuclear hormone receptor NHR-23 (also known as NR1F1) is an important regulator of C. elegans molting. NHR-23 expression oscillates in the epidermal epithelium, and soma-specific NHR-23 depletion causes severe developmental delay and death. Tissue-specific RNAi suggests that nhr-23 acts primarily in seam and hypodermal cells. NHR-23 coordinates the expression of factors involved in molting, lipid transport/metabolism and remodeling of the aECM. NHR-23 depletion causes dampened expression of a nas-37 promoter reporter and a loss of reporter oscillation. The cuticle collagen ROL-6 and zona pellucida protein NOAH-1 display aberrant annular localization and severe disorganization over the seam cells after NHR-23 depletion, while the expression of the adult-specific cuticle collagen BLI-1 is diminished and frequently found in patches. Consistent with these localization defects, the cuticle barrier is severely compromised when NHR-23 is depleted. Together, this work provides insight into how NHR-23 acts in the seam and hypodermal cells to coordinate aECM regeneration during development.
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- 2023
28. Experimental considerations for study of C. elegans lysosomal proteins
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Clancy, John C, Vo, An A, Myles, Krista M, Levenson, Max T, Ragle, James Matthew, and Ward, Jordan D
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Genetics ,Prevention ,Neurosciences ,Generic health relevance ,Animals ,Caenorhabditis elegans Proteins ,Caenorhabditis elegans ,Lysosomes ,Green Fluorescent Proteins ,lysosome ,Gamillus ,western blot ,Caenorhabditis elegans - Abstract
Lysosomes are an important organelle required for the degradation of a range of cellular components. Lysosome function is critical for development and homeostasis as dysfunction can lead to inherited genetic disorders, cancer, and neurodegenerative and metabolic diseases. The acidic and protease-rich environment of lysosomes poses experimental challenges. Many fluorescent proteins are quenched or degraded, while specific red fluorescent proteins can be cleaved from translational fusion partners and accumulate. While studying MLT-11, a Caenorhabditis elegans molting factor that localizes to lysosomes and the cuticle, we sought to optimize several experimental parameters. We found that, in contrast to mNeonGreen fusions, mScarlet fusions to MLT-11 missed cuticular and rectal epithelial localization. Rapid sample lysis and denaturation were critical for preventing MLT-11 fragmentation while preparing lysates for western blots. Using a model lysosomal substrate (NUC-1), we found that rigid polyproline linkers and truncated mCherry constructs do not prevent cleavage of mCherry from NUC-1. We provide evidence that extended localization in lysosomal environments prevents the detection of FLAG epitopes in western blots. Finally, we optimize an acid-tolerant green fluorescent protein (Gamillus) for use in C. elegans. These experiments provide important experimental considerations and new reagents for the study of C. elegans lysosomal proteins.
- Published
- 2023
29. Caenorhabditis elegans spermatocytes can segregate achiasmate homologous chromosomes apart at higher than random frequency during meiosis I
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Gong, Ting and McNally, Francis J
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Biochemistry and Cell Biology ,Genetics ,Biological Sciences ,Contraception/Reproduction ,1.1 Normal biological development and functioning ,Underpinning research ,Generic health relevance ,Animals ,Male ,Female ,Caenorhabditis elegans ,Spermatocytes ,Chromosomes ,Meiosis ,Caenorhabditis elegans Proteins ,Chromosome Segregation ,Aneuploidy ,meiosis ,chromosome segregation ,Developmental Biology ,Biochemistry and cell biology - Abstract
Chromosome segregation errors during meiosis are the leading cause of aneuploidy. Faithful chromosome segregation during meiosis in most eukaryotes requires a crossover which provides a physical attachment holding homologs together in a "bivalent." Crossovers are critical for homologs to be properly aligned and partitioned in the first meiotic division. Without a crossover, individual homologs (univalents) might segregate randomly, resulting in aneuploid progeny. However, Caenorhabditis elegans zim-2 mutants, which have crossover defects on chromosome V, have fewer dead embryos than that expected from random segregation. This deviation from random segregation is more pronounced in zim-2 males than that in females. We found three phenomena that can explain this apparent discrepancy. First, we detected crossovers on chromosome V in both zim-2(tm574) oocytes and spermatocytes, suggesting a redundant mechanism to make up for the ZIM-2 loss. Second, after accounting for the background crossover frequency, spermatocytes produced significantly more euploid gametes than what would be expected from random segregation. Lastly, trisomy of chromosome V is viable and fertile. Together, these three phenomena allow zim-2(tm574) mutants with reduced crossovers on chromosome V to have more viable progeny. Furthermore, live imaging of meiosis in spo-11(me44) oocytes and spermatocytes, which exhibit crossover failure on all 6 chromosomes, showed 12 univalents segregating apart in roughly equal masses in a homology-independent manner, supporting the existence of a mechanism that segregates any 2 chromosomes apart.
- Published
- 2023
30. ATM signaling modulates cohesin behavior in meiotic prophase and proliferating cells
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Yu, Zhouliang, Kim, Hyung Jun, and Dernburg, Abby F
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Biochemistry and Cell Biology ,Biomedical and Clinical Sciences ,Biological Sciences ,Genetics ,Underpinning research ,1.1 Normal biological development and functioning ,Generic health relevance ,Animals ,Meiosis ,Chromosomal Proteins ,Non-Histone ,Chromosomes ,Cell Cycle Proteins ,Chromosome Pairing ,Caenorhabditis elegans ,Mammals ,Chemical Sciences ,Medical and Health Sciences ,Biophysics ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences ,Chemical sciences - Abstract
Cohesins are ancient and ubiquitous regulators of chromosome architecture and function, but their diverse roles and regulation remain poorly understood. During meiosis, chromosomes are reorganized as linear arrays of chromatin loops around a cohesin axis. This unique organization underlies homolog pairing, synapsis, double-stranded break induction, and recombination. We report that axis assembly in Caenorhabditis elegans is promoted by DNA-damage response (DDR) kinases that are activated at meiotic entry, even in the absence of DNA breaks. Downregulation of the cohesin-destabilizing factor WAPL-1 by ATM-1 promotes axis association of cohesins containing the meiotic kleisins COH-3 and COH-4. ECO-1 and PDS-5 also contribute to stabilizing axis-associated meiotic cohesins. Further, our data suggest that cohesin-enriched domains that promote DNA repair in mammalian cells also depend on WAPL inhibition by ATM. Thus, DDR and Wapl seem to play conserved roles in cohesin regulation in meiotic prophase and proliferating cells.
- Published
- 2023
31. Potassium-chelating drug sodium polystyrene sulfonate enhances lysosomal function and suppresses proteotoxicity
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Arputhasamy, Cyrene, Foulger, Anna C, Lucanic, Mark, Rane, Anand, Schmidt, Minna, Garrett, Theo, Broussalian, Michael, Battistoni, Elena, Brem, Rachel B, Lithgow, Gordon J, Chamoli, Manish, and Andersen, Julie K
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Biochemistry and Cell Biology ,Biological Sciences ,Aging ,5.1 Pharmaceuticals ,Development of treatments and therapeutic interventions ,Generic health relevance ,Animals ,Humans ,Potassium ,Caenorhabditis elegans ,Prospective Studies ,Lysosomes ,C. elegans ,Lysosome ,Autophagy ,Potassium restriction ,Lifespan ,Proteotoxicity ,SH-SY5Y neuronal cells ,Amyloid-beta ,Neurodegeneration ,Amyloid-β ,Genetics ,Clinical sciences - Abstract
Lysosomes are crucial for degradation and recycling of damaged proteins and cellular components. Therapeutic strategies enhancing lysosomal function are a promising approach for aging and age-related neurodegenerative diseases. Here, we show that an FDA approved drug sodium polystyrene sulfonate (SPS), used to reduce high blood potassium in humans, enhances lysosomal function both in C. elegans and in human neuronal cells. Enhanced lysosomal function following SPS treatment is accompanied by the suppression of proteotoxicity caused by expression of the neurotoxic peptides Aβ and TAU. Additionally, treatment with SPS imparts health benefits as it significantly increases lifespan in C. elegans. Overall our work supports the potential use of SPS as a prospective geroprotective intervention.
- Published
- 2023
32. Redundant microtubule crosslinkers prevent meiotic spindle bending to ensure diploid offspring in C. elegans
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Li, Wenzhe, Crellin, Helena A, Cheerambathur, Dhanya, and McNally, Francis J
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Biochemistry and Cell Biology ,Biological Sciences ,Genetics ,Underpinning research ,1.1 Normal biological development and functioning ,Generic health relevance ,Animals ,Female ,Caenorhabditis elegans ,Kinesins ,Diploidy ,Caenorhabditis elegans Proteins ,Microtubules ,Spindle Apparatus ,Meiosis ,Oocytes ,Developmental Biology - Abstract
Oocyte meiotic spindles mediate the expulsion of ¾ of the genome into polar bodies to generate diploid zygotes in nearly all animal species. Failures in this process result in aneuploid or polyploid offspring that are typically inviable. Accurate meiotic chromosome segregation and polar body extrusion require the spindle to elongate while maintaining its structural integrity. Previous studies have implicated three hypothetical activities during this process, including microtubule crosslinking, microtubule sliding and microtubule polymerization. However, how these activities regulate spindle rigidity and elongation as well as the exact proteins involved in the activities remain unclear. We discovered that C. elegans meiotic anaphase spindle integrity is maintained through redundant microtubule crosslinking activities of the Kinesin-5 family motor BMK-1, the microtubule bundling protein SPD-1/PRC1, and the Kinesin-4 family motor, KLP-19. Using time-lapse imaging, we found that single depletion of KLP-19KIF4A, SPD-1PRC1 or BMK-1Eg5 had minimal effects on anaphase B spindle elongation velocity. In contrast, double depletion of SPD-1PRC1 and BMK-1Eg5 or double depletion of KLP-19KIF4A and BMK-1Eg5 resulted in spindles that elongated faster, bent in a myosin-dependent manner, and had a high rate of polar body extrusion errors. Bending spindles frequently extruded both sets of segregating chromosomes into two separate polar bodies. Normal anaphase B velocity was observed after double depletion of KLP-19KIF4A and SPD-1PRC1. These results suggest that KLP-19KIF4A and SPD-1PRC1 act in different pathways, each redundant with a separate BMK-1Eg5 pathway in regulating meiotic spindle elongation. Depletion of ZYG-8, a doublecortin-related microtubule binding protein, led to slower anaphase B spindle elongation. We found that ZYG-8DCLK1 acts by excluding SPD-1PRC1 from the spindle. Thus, three mechanistically distinct microtubule regulation modules, two based on crosslinking, and one based on exclusion of crosslinkers, power the mechanism that drives spindle elongation and structural integrity during anaphase B of C.elegans female meiosis.
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- 2023
33. Caenorhabditis elegans models for striated muscle disorders caused by missense variants of human LMNA
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Gregory, Ellen F, Kalra, Shilpi, Brock, Trisha, Bonne, Gisèle, Luxton, GW Gant, Hopkins, Christopher, and Starr, Daniel A
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Biological Sciences ,Genetics ,Heart Disease ,Cardiovascular ,Aetiology ,2.1 Biological and endogenous factors ,Animals ,Humans ,Caenorhabditis elegans ,Lamin Type A ,Laminopathies ,Muscle ,Skeletal ,Muscle ,Striated ,Muscular Diseases ,Mutation ,Missense ,Developmental Biology - Abstract
Striated muscle laminopathies caused by missense mutations in the nuclear lamin gene LMNA are characterized by cardiac dysfunction and often skeletal muscle defects. Attempts to predict which LMNA variants are pathogenic and to understand their physiological effects lag behind variant discovery. We created Caenorhabditis elegans models for striated muscle laminopathies by introducing pathogenic human LMNA variants and variants of unknown significance at conserved residues within the lmn-1 gene. Severe missense variants reduced fertility and/or motility in C. elegans. Nuclear morphology defects were evident in the hypodermal nuclei of many lamin variant strains, indicating a loss of nuclear envelope integrity. Phenotypic severity varied within the two classes of missense mutations involved in striated muscle disease, but overall, variants associated with both skeletal and cardiac muscle defects in humans lead to more severe phenotypes in our model than variants predicted to disrupt cardiac function alone. We also identified a separation of function allele, lmn-1(R204W), that exhibited normal viability and swimming behavior but had a severe nuclear migration defect. Thus, we established C. elegans avatars for striated muscle laminopathies and identified LMNA variants that offer insight into lamin mechanisms during normal development.
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- 2023
34. GPCR signaling regulates severe stress‐induced organismic death in Caenorhabditis elegans
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Wang, Changnan, Long, Yong, Wang, Bingying, Zhang, Chao, and K., Dengke
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Biochemistry and Cell Biology ,Biological Sciences ,Genetics ,Underpinning research ,1.1 Normal biological development and functioning ,Humans ,Animals ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Aging ,Basic-Leucine Zipper Transcription Factors ,Medical and Health Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences - Abstract
How an organism dies is a fundamental yet poorly understood question in biology. An organism can die of many causes, including stress-induced phenoptosis, also defined as organismic death that is regulated by its genome-encoded programs. The mechanism of stress-induced phenoptosis is still largely unknown. Here, we show that transient but severe freezing-thaw stress (FTS) in Caenorhabditis elegans induces rapid and robust phenoptosis that is regulated by G-protein coupled receptor (GPCR) signaling. RNAi screens identify the GPCR-encoding fshr-1 in mediating transcriptional responses to FTS. FSHR-1 increases ligand interaction upon FTS and activates a cyclic AMP-PKA cascade leading to a genetic program to promote organismic death under severe stress. FSHR-1/GPCR signaling up-regulates the bZIP-type transcription factor ZIP-10, linking FTS to expression of genes involved in lipid remodeling, proteostasis, and aging. A mathematical model suggests how genes may promote organismic death under severe stress conditions, potentially benefiting growth of the clonal population with individuals less stressed and more reproductively privileged. Our studies reveal the roles of FSHR-1/GPCR-mediated signaling in stress-induced gene expression and phenoptosis in C. elegans, providing empirical new insights into mechanisms of stress-induced phenoptosis with evolutionary implications.
- Published
- 2023
35. Differential requirement for BRCA1-BARD1 E3 ubiquitin ligase activity in DNA damage repair and meiosis in the Caenorhabditis elegans germ line
- Author
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Li, Qianyan, Kaur, Arshdeep, Okada, Kyoko, McKenney, Richard J, and Engebrecht, JoAnne
- Subjects
Biochemistry and Cell Biology ,Genetics ,Biological Sciences ,Cancer ,Underpinning research ,1.1 Normal biological development and functioning ,Generic health relevance ,Animals ,Caenorhabditis elegans ,Tumor Suppressor Proteins ,BRCA1 Protein ,Ubiquitin-Protein Ligases ,DNA Repair ,DNA Damage ,Meiosis ,Germ Cells ,Developmental Biology - Abstract
The tumor suppressor BRCA1-BARD1 complex regulates many cellular processes; of critical importance to its tumor suppressor function is its role in genome integrity. Although RING E3 ubiquitin ligase activity is the only known enzymatic activity of the complex, the in vivo requirement for BRCA1-BARD1 E3 ubiquitin ligase activity has been controversial. Here we probe the role of BRCA1-BARD1 E3 ubiquitin ligase activity in vivo using C. elegans. Genetic, cell biological, and biochemical analyses of mutants defective for E3 ligase activity suggest there is both E3 ligase-dependent and independent functions of the complex in the context of DNA damage repair and meiosis. We show that E3 ligase activity is important for nuclear accumulation of the complex and specifically to concentrate at meiotic recombination sites but not at DNA damage sites in proliferating germ cells. While BRCA1 alone is capable of monoubiquitylation, BARD1 is required with BRCA1 to promote polyubiquitylation. We find that the requirement for E3 ligase activity and BARD1 in DNA damage signaling and repair can be partially alleviated by driving the nuclear accumulation and self-association of BRCA1. Our data suggest that in addition to E3 ligase activity, BRCA1 may serve a structural role for DNA damage signaling and repair while BARD1 plays an accessory role to enhance BRCA1 function.
- Published
- 2023
36. Molecular evidence of widespread benzimidazole drug resistance in Ancylostoma caninum from domestic dogs throughout the USA and discovery of a novel β-tubulin benzimidazole resistance mutation
- Author
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Venkatesan, Abhinaya, Castro, Pablo D Jimenez, Morosetti, Arianna, Horvath, Hannah, Chen, Rebecca, Redman, Elizabeth, Dunn, Kayla, Collins, James Bryant, Fraser, James S, Andersen, Erik C, Kaplan, Ray M, and Gilleard, John S
- Subjects
Digestive Diseases ,Rare Diseases ,Emerging Infectious Diseases ,Vaccine Related ,Antimicrobial Resistance ,Genetics ,2.1 Biological and endogenous factors ,Aetiology ,Infection ,Animals ,Dogs ,Ancylostoma ,Ancylostomatoidea ,Anthelmintics ,Benzimidazoles ,Caenorhabditis elegans ,Drug Resistance ,Mutation ,Tubulin ,Microbiology ,Immunology ,Medical Microbiology ,Virology - Abstract
Ancylostoma caninum is an important zoonotic gastrointestinal nematode of dogs worldwide and a close relative of human hookworms. We recently reported that racing greyhound dogs in the USA are infected with A. caninum that are commonly resistant to multiple anthelmintics. Benzimidazole resistance in A. caninum in greyhounds was associated with a high frequency of the canonical F167Y(TTC>TAC) isotype-1 β-tubulin mutation. In this work, we show that benzimidazole resistance is remarkably widespread in A. caninum from domestic dogs across the USA. First, we identified and showed the functional significance of a novel benzimidazole isotype-1 β-tubulin resistance mutation, Q134H(CAA>CAT). Several benzimidazole resistant A. caninum isolates from greyhounds with a low frequency of the F167Y(TTC>TAC) mutation had a high frequency of a Q134H(CAA>CAT) mutation not previously reported from any eukaryotic pathogen in the field. Structural modeling predicted that the Q134 residue is directly involved in benzimidazole drug binding and that the 134H substitution would significantly reduce binding affinity. Introduction of the Q134H substitution into the C. elegans β-tubulin gene ben-1, by CRISPR-Cas9 editing, conferred similar levels of resistance as a ben-1 null allele. Deep amplicon sequencing on A. caninum eggs from 685 hookworm positive pet dog fecal samples revealed that both mutations were widespread across the USA, with prevalences of 49.7% (overall mean frequency 54.0%) and 31.1% (overall mean frequency 16.4%) for F167Y(TTC>TAC) and Q134H(CAA>CAT), respectively. Canonical codon 198 and 200 benzimidazole resistance mutations were absent. The F167Y(TTC>TAC) mutation had a significantly higher prevalence and frequency in Western USA than in other regions, which we hypothesize is due to differences in refugia. This work has important implications for companion animal parasite control and the potential emergence of drug resistance in human hookworms.
- Published
- 2023
37. NHR-23 and SPE-44 regulate distinct sets of genes during Caenorhabditis elegans spermatogenesis
- Author
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Ragle, James Matthew, Morrison, Kayleigh N, Vo, An A, Johnson, Zoe E, Lopez, Javier Hernandez, Rechtsteiner, Andreas, Shakes, Diane C, and Ward, Jordan D
- Subjects
Contraception/Reproduction ,Genetics ,1.1 Normal biological development and functioning ,Underpinning research ,Generic health relevance ,Animals ,Female ,Male ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Mutation ,Sperm Motility ,Semen ,Spermatogenesis ,Transcription Factors ,spermatogenesis ,NHR-23 ,SPE-44 ,nuclear hormone receptor ,transcription factor ,gene regulation ,Caenorhabditis elegans - Abstract
Spermatogenesis is the process through which mature male gametes are formed and is necessary for the transmission of genetic information. While much work has established how sperm fate is promoted and maintained, less is known about how the sperm morphogenesis program is executed. We previously identified a novel role for the nuclear hormone receptor transcription factor, NHR-23, in promoting Caenorhabditis elegans spermatogenesis. The depletion of NHR-23 along with SPE-44, another transcription factor that promotes spermatogenesis, caused additive phenotypes. Through RNA-seq, we determined that NHR-23 and SPE-44 regulate distinct sets of genes. The depletion of both NHR-23 and SPE-44 produced yet another set of differentially regulated genes. NHR-23-regulated genes are enriched in phosphatases, consistent with the switch from genome quiescence to post-translational regulation in spermatids. In the parasitic nematode Ascaris suum, MFP1 and MFP2 control the polymerization of Major Sperm Protein, the molecule that drives sperm motility and serves as a signal to promote ovulation. NHR-23 and SPE-44 regulate several MFP2 paralogs, and NHR-23 depletion from the male germline caused defective localization of MSD/MFP1 and NSPH-2/MFP2. Although NHR-23 and SPE-44 do not transcriptionally regulate the casein kinase gene spe-6, a key regulator of sperm development, SPE-6 protein is lost following NHR-23+SPE-44 depletion. Together, these experiments provide the first mechanistic insight into how NHR-23 promotes spermatogenesis and an entry point to understanding the synthetic genetic interaction between nhr-23 and spe-44.
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- 2022
38. A conserved megaprotein-based molecular bridge critical for lipid trafficking and cold resilience.
- Author
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Wang, Changnan, Wang, Bingying, Pandey, Taruna, Long, Yong, Zhang, Jianxiu, Oh, Fiona, Sima, Jessica, Guo, Ruyin, Liu, Yun, Zhang, Chao, Mukherjee, Shaeri, Bassik, Michael, Lin, Weichun, Deng, Huichao, Vale, Goncalo, McDonald, Jeffrey G, Shen, Kang, and Ma, Dengke K
- Subjects
Cell Membrane ,Endoplasmic Reticulum ,Animals ,Zebrafish ,Mammals ,Humans ,Caenorhabditis elegans ,Phospholipids ,Genetics ,2.1 Biological and endogenous factors ,Aetiology ,Generic health relevance - Abstract
Cells adapt to cold by increasing levels of unsaturated phospholipids and membrane fluidity through conserved homeostatic mechanisms. Here we report an exceptionally large and evolutionarily conserved protein LPD-3 in C. elegans that mediates lipid trafficking to confer cold resilience. We identify lpd-3 mutants in a mutagenesis screen for genetic suppressors of the lipid desaturase FAT-7. LPD-3 bridges the endoplasmic reticulum (ER) and plasma membranes (PM), forming a structurally predicted hydrophobic tunnel for lipid trafficking. lpd-3 mutants exhibit abnormal phospholipid distribution, diminished FAT-7 abundance, organismic vulnerability to cold, and are rescued by Lecithin comprising unsaturated phospholipids. Deficient lpd-3 homologues in Zebrafish and mammalian cells cause defects similar to those observed in C. elegans. As mutations in BLTP1, the human orthologue of lpd-3, cause Alkuraya-Kucinskas syndrome, LPD-3 family proteins may serve as evolutionarily conserved highway bridges critical for ER-associated non-vesicular lipid trafficking and resilience to cold stress in eukaryotic cells.
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- 2022
39. The relationship between intraflagellar transport and upstream protein trafficking pathways and macrocyclic lactone resistance in Caenorhabditis elegans.
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Brinzer, Robert A, Winter, Alan D, and Page, Antony P
- Subjects
- *
CARRIER proteins , *CAENORHABDITIS elegans , *PROTEIN transport , *CILIARY body , *MOXIDECTIN , *GENETICS , *QUORUM sensing - Abstract
Parasitic nematodes are globally important and place a heavy disease burden on infected humans, crops, and livestock, while commonly administered anthelmintics used for treatment are being rendered ineffective by increasing levels of resistance. It has recently been shown in the model nematode Caenorhabditis elegans that the sensory cilia of the amphid neurons play an important role in resistance toward macrocyclic lactones such as ivermectin (an avermectin) and moxidectin (a milbemycin) either through reduced uptake or intertissue signaling pathways. This study interrogated the extent to which ciliary defects relate to macrocyclic lactone resistance and dye-filling defects using a combination of forward genetics and targeted resistance screening approaches and confirmed the importance of intraflagellar transport in this process. This approach also identified the protein trafficking pathways used by the downstream effectors and the components of the ciliary basal body that are required for effector entry into these nonmotile structures. In total, 24 novel C. elegans anthelmintic survival-associated genes were identified in this study. When combined with previously known resistance genes, there are now 46 resistance-associated genes that are directly involved in amphid, cilia, and intraflagellar transport function. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
40. The effect of developmental variation on expression QTLs in a multi parental Caenorhabditis elegans population.
- Author
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Eijnatten, Abraham L van, Sterken, Mark G, Kammenga, Jan E, Nijveen, Harm, and Snoek, Basten L
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GENE expression , *CAENORHABDITIS elegans , *CAENORHABDITIS , *GENETIC regulation , *LOCUS (Genetics) , *AGE , *AMBIENT intelligence , *TEXTURE mapping - Abstract
Regulation of gene expression plays a crucial role in developmental processes and adaptation to changing environments. expression quantitative trait locus (eQTL) mapping is a technique used to study the genetic regulation of gene expression using the transcriptomes of recombinant inbred lines (RILs). Typically, the age of the inbred lines at the time of RNA sampling is carefully controlled. This is necessary because the developmental process causes changes in gene expression, complicating the interpretation of eQTL mapping experiments. However, due to genetics and variation in ambient micro-environments, organisms can differ in their "developmental age," even if they are of the same chronological age. As a result, eQTL patterns are affected by developmental variation in gene expression. The model organism Caenorhabditis elegans is particularly suited for studying the effect of developmental variation on eQTL mapping patterns. In a span of days, C. elegans transitions from embryo through 4 larval stages to adult while undergoing massive changes to its transcriptome. Here, we use C. elegans to investigate the effect of developmental age variation on eQTL patterns and present a normalization procedure. We used dynamical eQTL mapping, which includes the developmental age as a cofactor, to separate the variation in development from genotypic variation and explain variation in gene expression levels. We compare classical single marker eQTL mapping and dynamical eQTL mapping using RNA-seq data of ∼200 multi-parental RILs of C. elegans. The results show that (1) many eQTLs are caused by developmental variation, (2) most trans-bands are developmental QTLs, and (3) dynamical eQTL mapping detects additional eQTLs not found with classical eQTL mapping. We recommend that correction for variation in developmental age should be strongly considered in eQTL mapping studies given the large impact of processes like development on the transcriptome. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
41. Sperm-inherited H3K27me3 epialleles are transmitted transgenerationally in cis
- Author
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Kaneshiro, Kiyomi Raye, Egelhofer, Thea A, Rechtsteiner, Andreas, Cockrum, Chad, and Strome, Susan
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Reproductive Medicine ,Biochemistry and Cell Biology ,Biomedical and Clinical Sciences ,Biological Sciences ,Genetics ,Contraception/Reproduction ,1.1 Normal biological development and functioning ,Underpinning research ,Alleles ,Animals ,Caenorhabditis elegans ,Chromatin ,Epigenesis ,Genetic ,Histones ,Male ,Oocytes ,Semen ,Spermatozoa ,epigenetic inheritance ,gene regulation ,H3K27me3 ,C. elegans ,transgenerational - Abstract
The transmission of chromatin states from parent cells to daughter cells preserves cell-specific transcriptional states and thus cell identity through cell division. The mechanism that underpins this process is not fully understood. The role that chromatin states serve in transmitting gene expression information across generations via sperm and oocytes is even less understood. Here, we utilized a model in which Caenorhabditis elegans sperm and oocyte alleles were inherited in different states of the repressive mark H3K27me3. This resulted in the alleles achieving different transcriptional states within the nuclei of offspring. Using this model, we showed that sperm alleles inherited without H3K27me3 were sensitive to up-regulation in offspring somatic and germline tissues, and tissue context determined which genes were up-regulated. We found that the subset of sperm alleles that were up-regulated in offspring germlines retained the H3K27me3(-) state and were transmitted to grandoffspring as H3K27me3(-) and up-regulated epialleles, demonstrating that H3K27me3 can serve as a transgenerational epigenetic carrier in C. elegans.
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- 2022
42. The mIAA7 degron improves auxin-mediated degradation in Caenorhabditiselegans
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Sepers, Jorian J, Verstappen, Noud HM, Vo, An A, Ragle, James Matthew, Ruijtenberg, Suzan, Ward, Jordan D, and Boxem, Mike
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Bioengineering ,Biotechnology ,Generic health relevance ,Animals ,Caenorhabditis elegans ,Humans ,Indoleacetic Acids ,Nuclear Proteins ,Proteolysis ,mIAA7 ,TIR1 ,AID system ,protein degradation ,C aenorhabditis elegans ,C aenorhabditis elegans ,Genetics - Abstract
Auxin-inducible degradation is a powerful tool for the targeted degradation of proteins with spatiotemporal control. One limitation of the auxin-inducible degradation system is that not all proteins are degraded efficiently. Here, we demonstrate that an alternative degron sequence, termed mIAA7, improves the efficiency of degradation in Caenorhabditiselegans, as previously reported in human cells. We tested the depletion of a series of proteins with various subcellular localizations in different tissue types and found that the use of the mIAA7 degron resulted in faster depletion kinetics for 5 out of 6 proteins tested. The exception was the nuclear protein HIS-72, which was depleted with similar efficiency as with the conventional AID* degron sequence. The mIAA7 degron also increased the leaky degradation for 2 of the tested proteins. To overcome this problem, we combined the mIAA7 degron with the C. elegans AID2 system, which resulted in complete protein depletion without detectable leaky degradation. Finally, we show that the degradation of ERM-1, a highly stable protein that is challenging to deplete, could be improved further by using multiple mIAA7 degrons. Taken together, the mIAA7 degron further increases the power and applicability of the auxin-inducible degradation system. To facilitate the generation of mIAA7-tagged proteins using CRISPR/Cas9 genome engineering, we generated a toolkit of plasmids for the generation of dsDNA repair templates by PCR.
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- 2022
43. The X chromosome in C. elegans sex determination and dosage compensation
- Author
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Meyer, Barbara J
- Subjects
Biological Sciences ,Genetics ,Underpinning research ,1.1 Normal biological development and functioning ,Animals ,Caenorhabditis elegans ,Dosage Compensation ,Genetic ,Genes ,X-Linked ,Sex Chromosomes ,X Chromosome ,Developmental Biology ,Biochemistry and cell biology - Abstract
Abnormalities in chromosome dose can reduce organismal fitness and viability by disrupting the balance of gene expression. Unlike imbalances in chromosome dose that cause pathologies, differences in X-chromosome dose that determine sex are well tolerated. Dosage compensation mechanisms have evolved in diverse species to balance X-chromosome gene expression between sexes. Mechanisms underlying nematode X-chromosome counting to determine sex revealed how small quantitative differences in molecular signals are translated into dramatically different developmental fates. Mechanisms underlying X-chromosome dosage compensation revealed the interplay between chromatin modification and three-dimensional chromosome structure imposed by an X-specific condensin complex to regulate gene expression over vast chromosomal territories. In a surprising twist of evolution, this dosage-compensation condensin complex also regulates lifespan and tolerance to proteotoxic stress.
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- 2022
44. MEL-28/ELYS and CENP-C coordinately control outer kinetochore assembly and meiotic chromosome-microtubule interactions
- Author
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Hattersley, Neil, Schlientz, Aleesa J, Prevo, Bram, Oegema, Karen, and Desai, Arshad
- Subjects
Reproductive Medicine ,Biomedical and Clinical Sciences ,Biological Sciences ,Genetics ,1.1 Normal biological development and functioning ,Underpinning research ,Generic health relevance ,Animals ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Chromatin ,Chromosomal Proteins ,Non-Histone ,Chromosome Segregation ,DNA-Binding Proteins ,Kinetochores ,Microtubule-Associated Proteins ,Microtubules ,Mitosis ,Nuclear Pore Complex Proteins ,Spindle Apparatus ,C. elegans ,CENPA ,CENPC ,ELYS ,KMN network ,NDC80 ,centromere ,kinetochore ,meiosis ,nuclear pore ,oocyte ,spindle ,Medical and Health Sciences ,Psychology and Cognitive Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences ,Psychology - Abstract
During mitosis and meiosis in the majority of eukaryotes, centromeric chromatin comprised of CENP-A nucleosomes and their reader CENP-C recruits components of the outer kinetochore to build an interface with spindle microtubules.1,2 One exception is C. elegans oocyte meiosis, where outer kinetochore proteins form cup-like structures on chromosomes independently of centromeric chromatin.3 Here, we show that the nucleoporin MEL-28 (ortholog of human ELYS) and CENP-CHCP-4 act in parallel to recruit outer kinetochore components to oocyte meiotic chromosomes. Unexpectedly, co-inhibition of MEL-28 and CENP-CHCP-4 resulted in chromosomes being expelled from the meiotic spindle prior to anaphase onset, a more severe phenotype than what was observed following ablation of the outer kinetochore.4,5 This observation suggested that MEL-28 and the outer kinetochore independently link chromosomes to spindle microtubules. Consistent with this, the chromosome expulsion defect was observed following co-inhibition of MEL-28 and the microtubule-coupling KNL-1/MIS-12/NDC-80 (KMN) network of the outer kinetochore. Use of engineered mutants showed that MEL-28 acts in conjunction with the microtubule-binding NDC-80 complex to keep chromosomes within the oocyte meiotic spindle and that this function likely involves the Y-complex of nucleoporins that associate with MEL-28; by contrast, the ability to dock protein phosphatase 1, shared by MEL-28 and KNL-1, is not involved. These results highlight nuclear pore-independent functions for a conserved nucleoporin and explain two unusual features of oocyte meiotic chromosome segregation in C. elegans: centromeric chromatin-independent outer kinetochore assembly, and dispensability of the outer kinetochore for constraining chromosomes in the acentrosomal meiotic spindle.
- Published
- 2022
45. Inducible degradation of dosage compensation protein DPY-27 facilitates isolation of Caenorhabditis elegans males for molecular and biochemical analyses
- Author
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Li, Qianyan, Kaur, Arshdeep, Mallory, Benjamin, Hariri, Sara, and Engebrecht, JoAnne
- Subjects
Genetics ,Generic health relevance ,Animals ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Disorders of Sex Development ,Dosage Compensation ,Genetic ,Female ,Humans ,Male ,Meiosis ,X Chromosome ,dosage compensation ,DPY-27 ,males ,meiosis ,spermiogenesis ,Genetics of Sex ,Caenorhabditis elegans - Abstract
Biological sex affects numerous aspects of biology, yet how sex influences different biological processes have not been extensively studied at the molecular level. Caenorhabditis elegans, with both hermaphrodites (functionally females as adults) and males, is an excellent system to uncover how sex influences physiology. Here, we describe a method to isolate large quantities of C. elegans males by conditionally degrading DPY-27, a component of the dosage compensation complex essential for hermaphrodite, but not male, development. We show that germ cells from males isolated following DPY-27 degradation undergo meiosis and spermiogenesis like wild type and these males are competent to mate and sire viable offspring. We further demonstrate the efficacy of this system by analyzing gene expression and performing affinity pull-downs from male worm extracts.
- Published
- 2022
46. Transmembrane protein 120A (TMEM-120A/TACAN) coordinates with PIEZO channel during Caenorhabditis elegans reproductive regulation.
- Author
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Bai, Xiaofei and Golden, Andy
- Subjects
- *
MEMBRANE proteins , *CAENORHABDITIS elegans , *GENITALIA , *ANIMAL clutches , *PHYSIOLOGY , *ION channels - Abstract
Membrane protein TMEM120A (also known as TACAN) was presumed to be both a mechanically activated molecule and a lipid-modifying enzyme. TMEM120A has been identified as a negative regulator of the essential excitatory mechanosensitive protein PIEZO2. However, the extent to which TMEM120A mediates PIEZO2's activity during physiological processes remains largely unknown. In this study, we used the Caenorhabditis elegans reproductive tract to explore the functional contribution of tmem-120 , the sole TMEM120A/B ortholog, and its genetic interaction with pezo-1 in vivo. tmem-120 was expressed throughout the C. elegans development, particularly in the germline, embryos, and spermatheca. A tmem-120 mutant with a full-length deletion (tmem-120 Δ) displayed deformed germline, maternal sterility, and a reduced brood size. In vivo live imaging revealed that pinched zygotes were frequently observed in the uterus of tmem-120 Δ mutant animals, suggesting damage during spermathecal contraction. We then employed the auxin-inducible degradation system to degrade TMEM-120 protein in all somatic tissues or the germline, both of which resulted in reduced brood sizes. These findings suggested that multiple inputs of tmem-120 from different tissues regulate reproduction. Lastly, the loss of tmem-120 alleviated the brood size reduction and defective sperm navigation behavior in the pezo-1 Δ mutant. Overall, our findings reveal a role for tmem-120 in regulating reproductive physiology in C. elegans , and suggest an epistatic interaction between pezo-1 and tmem-120 when governing proper reproduction. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
47. Orthologs of the Caenorhabditis elegans heterochronic genes have divergent functions in Caenorhabditis briggsae.
- Author
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Ivanova, Maria and Moss, Eric G.
- Subjects
- *
GENETICS , *NEMATODES , *GENETIC mutation , *SEQUENCE analysis , *CAENORHABDITIS elegans , *MICROSCOPY , *SPRAINS , *GENES , *DESCRIPTIVE statistics , *CRISPRS , *PHENOTYPES - Abstract
The heterochronic genes of Caenorhabditis elegans comprise the best-studied pathway controlling the timing of tissue and organ formation in an animal. To begin to understand the evolution of this pathway and the significance of the relationships among its components, we characterized 11 Caenorhabditis briggsae orthologs of C. elegans heterochronic genes. Using CRISPR/Cas9, we made a variety of alleles and found that several mutant phenotypes differ in significant ways from those of C. elegans. Although most mutant orthologs displayed defects in developmental timing, their phenotypes could differ in which stages were affected, the penetrance and expressivity of the phenotypes, or by having additional pleiotropies that were not obviously connected to developmental timing. However, when examining pairwise epistasis and synergistic relationships, we found those paralleled the known relationships between their C. elegans orthologs, suggesting that the arrangements of these genes in functional modules are conserved, but the modules’ relationships to each other and/or to their targets has drifted since the time of the species’ last common ancestor. Furthermore, our investigation has revealed a relationship between this pathway to other aspects of the animal’s growth and development, including gonad development, which is relevant to both species. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
48. Correction: It's All in Your Mind: Determining Germ Cell Fate by Neuronal IRE-1 in C. elegans.
- Author
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Levi-Ferber, Mor, Salzberg, Yehuda, Safra, Modi, Haviv-Chesner, Anat, Bülow, Hannes E., and Henis-Korenblit, Sivan
- Subjects
- *
CAENORHABDITIS elegans , *GERM cells , *GENETICS - Abstract
This document is a correction notice for an article published in PLoS Genetics. Concerns were raised about several figures in the article, including duplication of panels and incorrect labeling. The corresponding author provided updated versions of the figures and underlying data from replicate experiments. An expert reviewer confirmed that the conclusions of the article are still supported. The primary data underlying the article are no longer available, except for some data underlying one figure. [Extracted from the article]
- Published
- 2023
- Full Text
- View/download PDF
49. WormBase in 2022—data, processes, and tools for analyzing Caenorhabditis elegans
- Author
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Davis, Paul, Zarowiecki, Magdalena, Arnaboldi, Valerio, Becerra, Andrés, Cain, Scott, Chan, Juancarlos, Chen, Wen J, Cho, Jaehyoung, da Veiga Beltrame, Eduardo, Diamantakis, Stavros, Gao, Sibyl, Grigoriadis, Dionysis, Grove, Christian A, Harris, Todd W, Kishore, Ranjana, Le, Tuan, Lee, Raymond YN, Luypaert, Manuel, Müller, Hans-Michael, Nakamura, Cecilia, Nuin, Paulo, Paulini, Michael, Quinton-Tulloch, Mark, Raciti, Daniela, Rodgers, Faye H, Russell, Matthew, Schindelman, Gary, Singh, Archana, Stickland, Tim, Van Auken, Kimberly, Wang, Qinghua, Williams, Gary, Wright, Adam J, Yook, Karen, Berriman, Matt, Howe, Kevin L, Schedl, Tim, Stein, Lincoln, and Sternberg, Paul W
- Subjects
Biological Sciences ,Genetics ,Human Genome ,Generic health relevance ,Animals ,Caenorhabditis ,Caenorhabditis elegans ,Databases ,Genetic ,Genome ,Genomics ,Humans ,Nematoda ,Caenorhabditis elegans ,annotation ,caenorhabditis ,community ,curation ,data ,database ,gene ,health ,human ,literature ,mining ,model ,nematode ,platform ,research ,resource ,software ,tools ,Developmental Biology ,Biochemistry and cell biology - Abstract
WormBase (www.wormbase.org) is the central repository for the genetics and genomics of the nematode Caenorhabditis elegans. We provide the research community with data and tools to facilitate the use of C. elegans and related nematodes as model organisms for studying human health, development, and many aspects of fundamental biology. Throughout our 22-year history, we have continued to evolve to reflect progress and innovation in the science and technologies involved in the study of C. elegans. We strive to incorporate new data types and richer data sets, and to provide integrated displays and services that avail the knowledge generated by the published nematode genetics literature. Here, we provide a broad overview of the current state of WormBase in terms of data type, curation workflows, analysis, and tools, including exciting new advances for analysis of single-cell data, text mining and visualization, and the new community collaboration forum. Concurrently, we continue the integration and harmonization of infrastructure, processes, and tools with the Alliance of Genome Resources, of which WormBase is a founding member.
- Published
- 2022
50. Harmonizing model organism data in the Alliance of Genome Resources
- Author
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Agapite, Julie, Albou, Laurent-Philippe, Aleksander, Suzanne A, Alexander, Micheal, Anagnostopoulos, Anna V, Antonazzo, Giulia, Argasinska, Joanna, Arnaboldi, Valerio, Attrill, Helen, Becerra, Andrés, Bello, Susan M, Blake, Judith A, Blodgett, Olin, Bradford, Yvonne M, Bult, Carol J, Cain, Scott, Calvi, Brian R, Carbon, Seth, Chan, Juancarlos, Chen, Wen J, Cherry, J Michael, Cho, Jaehyoung, Christie, Karen R, Crosby, Madeline A, Davis, Paul, da Veiga Beltrame, Eduardo, De Pons, Jeffrey L, D’Eustachio, Peter, Diamantakis, Stavros, Dolan, Mary E, dos Santos, Gilberto, Douglass, Eric, Dunn, Barbara, Eagle, Anne, Ebert, Dustin, Engel, Stacia R, Fashena, David, Foley, Saoirse, Frazer, Ken, Gao, Sibyl, Gibson, Adam C, Gondwe, Felix, Goodman, Josh, Gramates, L Sian, Grove, Christian A, Hale, Paul, Harris, Todd, Hayman, G Thomas, Hill, David P, Howe, Douglas G, Howe, Kevin L, Hu, Yanhui, Jha, Sagar, Kadin, James A, Kaufman, Thomas C, Kalita, Patrick, Karra, Kalpana, Kishore, Ranjana, Kwitek, Anne E, Laulederkind, Stanley JF, Lee, Raymond, Longden, Ian, Luypaert, Manuel, MacPherson, Kevin A, Martin, Ryan, Marygold, Steven J, Matthews, Beverley, McAndrews, Monica S, Millburn, Gillian, Miyasato, Stuart, Motenko, Howie, Moxon, Sierra, Muller, Hans-Michael, Mungall, Christopher J, Muruganujan, Anushya, Mushayahama, Tremayne, Nalabolu, Harika S, Nash, Robert S, Ng, Patrick, Nuin, Paulo, Paddock, Holly, Paulini, Michael, Perrimon, Norbert, Pich, Christian, Quinton-Tulloch, Mark, Raciti, Daniela, Ramachandran, Sridhar, Richardson, Joel E, Gelbart, Susan Russo, Ruzicka, Leyla, Schaper, Kevin, Schindelman, Gary, Shimoyama, Mary, Simison, Matt, Shaw, David R, Shrivatsav, Ajay, Singer, Amy, Skrzypek, Marek, Smith, Constance M, and Smith, Cynthia L
- Subjects
Biological Sciences ,Bioinformatics and Computational Biology ,Genetics ,Biotechnology ,Human Genome ,Underpinning research ,1.5 Resources and infrastructure (underpinning) ,Alleles ,Animals ,Caenorhabditis elegans ,Databases ,Genetic ,Drosophila ,Gene Ontology ,Humans ,Internet ,Mice ,Molecular Sequence Annotation ,Rats ,Saccharomycetales ,Zebrafish ,Alliance of Genome Resources Consortium ,biocuration ,data mining ,gene expression ,gene function ,gene interaction ,genome ,knowledgebase ,phenotype ,variants ,Developmental Biology ,Biochemistry and cell biology - Abstract
The Alliance of Genome Resources (the Alliance) is a combined effort of 7 knowledgebase projects: Saccharomyces Genome Database, WormBase, FlyBase, Mouse Genome Database, the Zebrafish Information Network, Rat Genome Database, and the Gene Ontology Resource. The Alliance seeks to provide several benefits: better service to the various communities served by these projects; a harmonized view of data for all biomedical researchers, bioinformaticians, clinicians, and students; and a more sustainable infrastructure. The Alliance has harmonized cross-organism data to provide useful comparative views of gene function, gene expression, and human disease relevance. The basis of the comparative views is shared calls of orthology relationships and the use of common ontologies. The key types of data are alleles and variants, gene function based on gene ontology annotations, phenotypes, association to human disease, gene expression, protein-protein and genetic interactions, and participation in pathways. The information is presented on uniform gene pages that allow facile summarization of information about each gene in each of the 7 organisms covered (budding yeast, roundworm Caenorhabditis elegans, fruit fly, house mouse, zebrafish, brown rat, and human). The harmonized knowledge is freely available on the alliancegenome.org portal, as downloadable files, and by APIs. We expect other existing and emerging knowledge bases to join in the effort to provide the union of useful data and features that each knowledge base currently provides.
- Published
- 2022
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