1,784 results
Search Results
2. Correction.
- Subjects
THYROID hormone receptors ,RNA sequencing ,GENE expression ,TEMPORAL lobe epilepsy ,NON-coding RNA - Abstract
This document is a correction notice for an article titled "A systems approach delivers a functional microRNA catalog and expanded targets for seizure suppression in temporal lobe epilepsy." The authors acknowledge an error in Figure 7D, where the x-axis was plotted incorrectly. However, this error does not affect the main results or conclusions of the paper, which identified microRNAs and the TGF-beta signaling pathway as potential therapeutic targets for epilepsy. The corrected figure and legend have been provided, and the raw data remains unchanged. The authors also note some corrections to the text in certain sections of the article. [Extracted from the article]
- Published
- 2024
- Full Text
- View/download PDF
3. Remethylation of Dnmt3a -/- hematopoietic cells is associated with partial correction of gene dysregulation and reduced myeloid skewing.
- Author
-
Ketkar S, Verdoni AM, Smith AM, Bangert CV, Leight ER, Chen DY, Brune MK, Helton NM, Hoock M, George DR, Fronick C, Fulton RS, Ramakrishnan SM, Chang GS, Petti AA, Spencer DH, Miller CA, and Ley TJ
- Subjects
- Animals, Bone Marrow Transplantation, DNA Methyltransferase 3A, Hematopoiesis genetics, Humans, Mice, Mice, Transgenic, Mutation genetics, Bone Marrow Cells metabolism, DNA (Cytosine-5-)-Methyltransferases genetics, DNA (Cytosine-5-)-Methyltransferases metabolism, DNA Methylation genetics, Gene Expression genetics
- Abstract
Mutations in the DNA methyltransferase 3A ( DNMT3A ) gene are the most common cause of age-related clonal hematopoiesis (ARCH) in older individuals, and are among the most common initiating events for acute myeloid leukemia (AML). The most frequent DNMT3A mutation in AML patients (R882H) encodes a dominant-negative protein that reduces methyltransferase activity by ∼80% in cells with heterozygous mutations, causing a focal, canonical DNA hypomethylation phenotype; this phenotype is partially recapitulated in murine Dnmt3a
-/- bone marrow cells. To determine whether the hypomethylation phenotype of Dnmt3a-/- hematopoietic cells is reversible, we developed an inducible transgene to restore expression of DNMT3A addback partially corrected dysregulated gene expression, and mitigated the expansion of myeloid cells. These data show that restoring Dnmt3a-/- mice. Partial remethylation was detected within 1 wk, but near-complete remethylation required 6 mo. Remethylation was accurate, dynamic, and highly ordered, suggesting that differentially methylated regions have unique properties that may be relevant for their functions. Importantly, 22 wk of DNMT3A addback partially corrected dysregulated gene expression, and mitigated the expansion of myeloid cells. These data show that restoring DNMT3A expression can alter the epigenetic "state" created by loss of Dnmt3a activity; this genetic proof-of-concept experiment suggests that this approach could be relevant for patients with ARCH or AML caused by loss-of-function DNMT3A mutations., Competing Interests: Competing interest statement: T.J.L. is engaged in a scientific collaboration with Rigel Pharmaceuticals to develop drugs that inhibit the function of the DNMT3A R882H mutation. He has received two honoraria from Rigel Pharmaceuticals to give presentations at their company in South San Francisco, CA. None of this work is described in this paper., (Copyright © 2020 the Author(s). Published by PNAS.)- Published
- 2020
- Full Text
- View/download PDF
4. Nrf2 activation attenuates genetic endoplasmic reticulum stress induced by a mutation in the phosphomannomutase 2 gene in zebrafish.
- Author
-
Katsuki Mukaigasa, Tadayuki Tsujita, Vu Thanh Nguyen, Li Li, Hirokazu Yagi, Yuji Fuse, Yaeko Nakajima-Takagi, Koichi Kato, Masayuki Yamamoto, and Makoto Kobayashi
- Subjects
ENDOPLASMIC reticulum ,PHOSPHOMANNOMUTASE ,SULFORAPHANE ,GENE expression ,PROTEIN kinases - Abstract
Nrf2 plays critical roles in animals' defense against electrophiles and oxidative stress by orchestrating the induction of cytoprotective genes. We previously isolated the zebrafish mutant it768, which displays up-regulated expression of Nrf2 target genes in an uninduced state. In this paper, we determine that the gene responsible for it768 was the zebrafish homolog of phosphomannomutase 2 (Pmm2), which is a key enzyme in the initial steps of N-glycosylation, and its mutation in humans leads to PMM2-CDG (congenital disorders of glycosylation), the most frequent type of CDG. The pmm2
it768 larvae exhibited mild defects in N-glycosylation, indicating that the pmm2it768 mutation is a hypomorph, as in human PMM2-CDG patients. A gene expression analysis showed that pmm2it768 larvae display up-regulation of endoplasmic reticulum (ER) stress, suggesting that the activation of Nrf2 was induced by the ER stress. Indeed, the treatment with the ER stress-inducing compounds up-regulated the gstpl expression in an Nrf2-dependent manner. Furthermore, the up-regulation of gstpl by the pmm2 inactivation was diminished by knocking down or out double-stranded RNA-activated protein kinase (PKR)-like ER kinase (PERK), one of the main ER stress sensors, suggesting that Nrf2 was activated in response to the ER stress via the PERK pathway. ER stress-induced activation of Nrf2 was reported previously, but the results have been controversial. Our present study clearly demonstrated that ER stress can indeed activate Nrf2 and this regulation is evolutionarily conserved among vertebrates. Moreover, ER stress induced in pmm2it768 mutants was ameliorated by the treatment of the Nrf2-activator sulforaphane, indicating that Nrf2 plays significant roles in the reduction of ER stress. [ABSTRACT FROM AUTHOR]- Published
- 2018
- Full Text
- View/download PDF
5. InfoBiology by printed arrays of microorganism colonies for timed and on-demand release of messages.
- Author
-
Palacios, Manuel A., Benito!Peña, Elena, Manesse, Mael, Mazzeo, Aaron D., LaFratta, Christopher N., Whitesides, George M., and Walt, David R.
- Subjects
BIOTECHNOLOGY ,INFORMATION technology ,BACTERIAL genetics ,ESCHERICHIA coli ,PHENOTYPES ,GENE expression - Abstract
This paper presents a proof-of-principle method, called InfoBiology, to write and encode data using arrays of genetically engineered strains of Escherichia coli with fluorescent proteins (FPs) as phenotypic markers. In InfoBiology, we encode, send, and release information using living organisms as carriers of data. Genetically engineered systems offer exquisite control of both genotype and phenotype. Living systems also offer the possibility for timed release of information as phenotypic features can take hours or days to develop. We use growth media and chemically induced gene expression as cipher keys or "biociphers" to develop encoded messages. The messages, called Steganography by Printed Arrays of Microbes (SPAM), consist of a matrix of spots generated by seven strains of E. coli, with each strain expressing a different FP. The coding scheme for these arrays relies on strings of paired, septenary digits, where each pair represents an alphanumeric character. In addition, the photophysical properties of the FPs offer another method for ciphering messages. Unique combinations of excited and emitted wavelengths generate distinct fluorescent patterns from the Steganography by Printed Arrays of Microbes (SPAM). This paper shows a new form of steganography based on information from engineered living systems. The combination of bio- and "photociphers" along with controlled timed-release exemplify the capabilities of InfoBiology, which could enable biometrics, communication through compromised channels, easy-to-read barcoding of biological products, or provide a deterrent to counterfeiting. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
6. Single-nuclei sequencing of uterine serous carcinoma reveals racial differences in immune signaling.
- Author
-
Foley, K. Grace, Adli, Mazhar, and Kim, J. Julie
- Subjects
BLACK people ,CELL populations ,GENE expression ,TUMOR microenvironment ,RACIAL inequality - Abstract
Significant racial disparities exist between Black and White patients with uterine serous carcinoma (USC). While the reasons for these disparities are unclear, several studies have demonstrated significantly different rates of driver mutations between racial groups, including TP53. However, limited research has investigated the transcriptional differences of tumors or the composition of the tumor microenvironment (TME) between these groups. Here, we report the single-nuclei RNA-sequencing profiles of primary USC tumors from diverse racial backgrounds. We find that there are significant differences between the tumors of Black and White patients. Tumors from Black patients exhibited higher expression of specific genes associated with aggressiveness, such as PAX8, and axon guidance and synaptic signaling pathways. We also demonstrated that T cell populations are reduced in the tumor tissue compared to matched benign, while anti-inflammatory macrophage populations are retained within the TME. Furthermore, we investigated the connection between PAX8 overexpression and immunosuppression in USC through regulation of several cytokines and chemokines. Notably, we show that PAX8 activity can influence macrophage gene expression and protein secretion. These studies provide a detailed understanding of the USC transcriptome and TME, and identify differences in tumor biology from patients of different racial backgrounds. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
7. The neocortical infrastructure for language involves region-specific patterns of laminar gene expression.
- Author
-
Wong, Maggie M. K., Zhiqiang Sha, Lütje, Lukas, Xiang-Zhen Kong, van Heukelum, Sabrina, van de Berg, Wilma D. J., Jonkman, Laura E., Fisher, Simon E., and Francks, Clyde
- Subjects
TEMPORAL lobe ,GENE expression ,FRONTAL lobe ,TRANSCRIPTOMES ,MEDICAL screening ,PEOPLE with schizophrenia - Abstract
The language network of the human brain has core components in the inferior frontal cortex and superior/middle temporal cortex, with left-hemisphere dominance in most people. Functional specialization and interconnectivity of these neocortical regions is likely to be reflected in their molecular and cellular profiles. Excitatory connections between cortical regions arise and innervate according to layer-specific patterns. Here, we generated a gene expression dataset from human postmortem cortical tissue samples from core language network regions, using spatial transcriptomics to discriminate gene expression across cortical layers. Integration of these data with existing single-cell expression data identified 56 genes that showed differences in laminar expression profiles between the frontal and temporal language cortex together with upregulation in layer II/III and/or layer V/VI excitatory neurons. Based on data from large-scale genome-wide screening in the population, DNA variants within these 56 genes showed set-level associations with interindividual variation in structural connectivity between the left-hemisphere frontal and temporal language cortex, and with the brain-related disorders dyslexia and schizophrenia which often involve affected language. These findings identify region-specific patterns of laminar gene expression as a feature of the brain's language network. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
8. Genetic mechanisms for impaired synaptic plasticity in schizophrenia revealed by computational modeling.
- Author
-
Mäki-Marttunen, Tuomo, Blackwell, Kim T., Akkouh, Ibrahim, Shadrin, Alexey, Valstad, Mathias, Elvsåshagen, Torbjørn, Linne, Marja-Leena, Djurovic, Srdjan, Einevoll, Gaute T., and Andreassen, Ole A.
- Subjects
GENE expression ,VISUAL evoked potentials ,LONG-term potentiation ,NEUROPLASTICITY ,CINGULATE cortex - Abstract
Schizophrenia phenotypes are suggestive of impaired cortical plasticity in the disease, but the mechanisms of these deficits are unknown. Genomic association studies have implicated a large number of genes that regulate neuromodulation and plasticity, indicating that the plasticity deficits have a genetic origin. Here, we used biochemically detailed computational modeling of postsynaptic plasticity to investigate how schizophrenia-associated genes regulate long-term potentiation (LTP) and depression (LTD). We combined our model with data from postmortem RNA expression studies (CommonMind gene-expression datasets) to assess the consequences of altered expression of plasticity-regulating genes for the amplitude of LTP and LTD. Our results show that the expression alterations observed post mortem, especially those in the anterior cingulate cortex, lead to impaired protein kinase A (PKA)-pathwaymediated LTP in synapses containing GluR1 receptors. We validated these findings using a genotyped electroencephalogram (EEG) dataset where polygenic risk scores for synaptic and ion channel-encoding genes as well as modulation of visual evoked potentials were determined for 286 healthy controls. Our results provide a possible genetic mechanism for plasticity impairments in schizophrenia, which can lead to improved understanding and, ultimately, treatment of the disorder. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
9. Leveraging a large language model to predict protein phase transition: A physical, multiscale, and interpretable approach.
- Author
-
Frank, Mor, Pengyu Ni, Jensen, Matthew, and Gerstein, Mark B.
- Subjects
LANGUAGE models ,PROTEIN structure prediction ,PHASE transitions ,ALZHEIMER'S disease ,PHASE separation - Abstract
Protein phase transitions (PPTs) from the soluble state to a dense liquid phase (forming droplets via liquid-liquid phase separation) or to solid aggregates (such as amyloids) play key roles in pathological processes associated with age-related diseases such as Alzheimer's disease. Several computational frameworks are capable of separately predicting the formation of droplets or amyloid aggregates based on protein sequences, yet none have tackled the prediction of both within a unified framework. Recently, large language models (LLMs) have exhibited great success in protein structure prediction; however, they have not yet been used for PPTs. Here, we fine-tune a LLM for predicting PPTs and demonstrate its usage in evaluating how sequence variants affect PPTs, an operation useful for protein design. In addition, we show its superior performance compared to suitable classical benchmarks. Due to the "black-box" nature of the LLM, we also employ a classical random forest model along with biophysical features to facilitate interpretation. Finally, focusing on Alzheimer's disease-related proteins, we demonstrate that greater aggregation is associated with reduced gene expression in Alzheimer's disease, suggesting a natural defense mechanism. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
10. Competing adaptations maintain nonadaptive variation in a wild cricket population.
- Author
-
Rayner, Jack G., Eichenberger, Franca, Bainbridge, Jessica V. A., Shangzhe Zhang, Xiao Zhang, Yusuf, Leeban H., Balenger, Susan, Gaggiotti, Oscar E., and Bailey, Nathan W.
- Subjects
BIOLOGICAL evolution ,GENE expression ,PHENOTYPIC plasticity ,PHENOTYPES ,POLYMORPHISM (Zoology) - Abstract
How emerging adaptive variants interact is an important factor in the evolution of wild populations, but the opportunity to empirically study this interaction is rare. We recently documented the emergence of an adaptive phenotype "curly-wing" in Hawaiian populations of field crickets (Teleogryllus oceanicus). Curly-wing inhibits males' ability to sing, protecting them from eavesdropping parasitoid flies (Ormia ochracea). Surprisingly, curly-wing co-occurs with similarly protective silent "flatwing" phenotypes in multiple populations, in which neither phenotype has spread to fixation. These two phenotypes are frequently coexpressed, but since either sufficiently reduces song amplitude to evade the fly, their coexpression confers no additional fitness benefit. Numerous "off-target" phenotypic changes are known to accompany flatwing, and we find that curly-wing, too, negatively impacts male courtship ability and affects mass and survival of females under lab conditions. We show through crosses and genomic and mRNA sequencing that curly-wing expression is associated with variation on a single autosome. In parallel analyses of flatwing, our results reinforce previous findings of X-linked single-locus inheritance. By combining insights into the genetic architecture of these alternative phenotypes with simulations and field observations, we show that the co-occurrence of these two adaptations impedes either from fixing, despite extreme fitness benefits, due to fitness epistasis. This co-occurrence of similar adaptive forms in the same populations might be more common than is generally considered and could be an important force inhibiting adaptive evolution in wild populations of sexually reproducing organisms. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
11. Unraveling clonal CD8 T cell expansion and identification of essential factors in γ-herpesvirus-induced lymphomagenesis.
- Author
-
Meijiao Gong, Myster, Françoise, Azouz, Abdulkader, Sanchez, Guillem Sanchez, Shifang Li, Charloteaux, Benoit, Bin Yang, Nichols, Jenna, Lefevre, Lucas, Javaux, Justine, Leemans, Sylvain, Nivelles, Olivier, van Campe, Willem, Roels, Stefan, Mostin, Laurent, van den Berg, Thierry, Davison, Andrew J., Gillet, Laurent, Connelley, Timothy, and Vermijlen, David
- Subjects
T cell receptors ,T cells ,MEMBRANE proteins ,CELLULAR signal transduction ,GENE expression - Abstract
Alcelaphine gammaherpesvirus 1 (AlHV-1) asymptomatically persists in its natural host, the wildebeest. However, cross-species transmission to cattle results in the induction of an acute and lethal peripheral T cell lymphoma-like disease (PTCL), named malignant catarrhal fever (MCF). Our previous findings demonstrated an essential role for viral genome maintenance in infected CD8+ T lymphocytes but the exact mechanism(s) leading to lymphoproliferation and MCF remained unknown. To decipher how AlHV-1 dysregulates T lymphocytes, we first examined the global phenotypic changes in circulating CD8+ T cells after experimental infection of calves. T cell receptor repertoire together with transcriptomics and epigenomics analyses demonstrated an oligoclonal expansion of infected CD8+ T cells displaying effector and exhaustion gene signatures, including GZMA, GNLY, PD-1, and TOX2 expression. Then, among viral genes expressed in infected CD8+ T cells, we uncovered A10 that encodes a transmembrane signaling protein displaying multiple tyrosine residues, with predicted ITAM and SH3 motifs. Impaired A10 expression did not affect AlHV-1 replication in vitro but rendered AlHV-1 unable to induce MCF. Furthermore, A10 was phosphorylated in T lymphocytes in vitro and affected T cell signaling. Finally, while AlHV-1 mutants expressing mutated forms of A10 devoid of ITAM or SH3 motifs (or both) were able to induce MCF, a recombinant virus expressing a mutated form of A10 unable to phosphorylate its tyrosine residues resulted in the lack of MCF and protected against a wild-type virus challenge. Thus, we could characterize the nature of this γ-herpesvirus-induced PTCL-like disease and identify an essential mechanism explaining its development. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
12. Matrix stiffness-dependent regulation of immunomodulatory genes in human MSCs is associated with the lncRNA CYTOR.
- Author
-
Lim, Justin J., Vining, Kyle H., Mooney, David J., and Blencowe, Benjamin J.
- Subjects
GENE expression ,LINCRNA ,CELL morphology ,STROMAL cells ,MECHANOTRANSDUCTION (Cytology) - Abstract
Cell-matrix interactions in 3D environments significantly differ from those in 2D cultures. As such, mechanisms of mechanotransduction in 2D cultures are not necessarily applicable to cell-encapsulating hydrogels that resemble features of tissue architecture. Accordingly, the characterization of molecular pathways in 3D matrices is expected to uncover insights into how cells respond to their mechanical environment in physiological contexts, and potentially also inform hydrogel-based strategies in cell therapies. In this study, a bone marrow-mimetic hydrogel was employed to systematically investigate the stiffness-responsive transcriptome of mesenchymal stromal cells. High matrix rigidity impeded integrin-collagen adhesion, resulting in changes in cell morphology characterized by a contractile network of actin proximal to the cell membrane. This resulted in a suppression of extracellular matrix-regulatory genes involved in the remodeling of collagen fibrils, as well as the upregulation of secreted immunomodulatory factors. Moreover, an investigation of long noncoding RNAs revealed that the cytoskeleton regulator RNA (CYTOR) contributes to these 3D stiffness-driven changes in gene expression. Knockdown of CYTOR using antisense oligonucleotides enhanced the expression of numerous mechanoresponsive cytokines and chemokines to levels exceeding those achievable by modulating matrix stiffness alone. Taken together, our findings further our understanding of mechanisms of mechanotransduction that are distinct from canonical mechanotransductive pathways observed in 2D cultures. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
13. Reconstruction of single-cell lineage trajectories and identification of diversity in fates during the epithelial-to-mesenchymal transition.
- Author
-
Yu-Chen Cheng, Yun Zhang, Tripathi, Shubham, Harshavardhan, B. V., Jolly, Mohit Kumar, Schiebinger, Geoffrey, Levine, Herbert, McDonald, Thomas O., and Michor, Franziska
- Subjects
EPITHELIAL-mesenchymal transition ,GENE expression ,RNA sequencing ,CELL cycle ,CELL lines - Abstract
Exploring the complexity of the epithelial-to-mesenchymal transition (EMT) unveils a diversity of potential cell fates; however, the exact timing and mechanisms by which early cell states diverge into distinct EMT trajectories remain unclear. Studying these EMT trajectories through single-cell RNA sequencing is challenging due to the necessity of sacrificing cells for each measurement. In this study, we employed optimal-transport analysis to reconstruct the past trajectories of different cell fates during TGF-beta-induced EMT in the MCF10A cell line. Our analysis revealed three distinct trajectories leading to low EMT, partial EMT, and high EMT states. Cells along the partial EMT trajectory showed substantial variations in the EMT signature and exhibited pronounced stemness. Throughout this EMT trajectory, we observed a consistent downregulation of the EED and EZH2 genes. This finding was validated by recent inhibitor screens of EMT regulators and CRISPR screen studies. Moreover, we applied our analysis of early-phase differential gene expression to gene sets associated with stemness and proliferation, pinpointing ITGB4, LAMA3, and LAMB3 as genes differentially expressed in the initial stages of the partial versus high EMT trajectories. We also found that CENPF, CKS1B, and MKI67 showed significant upregulation in the high EMT trajectory. While the first group of genes aligns with findings from previous studies, our work uniquely pinpoints the precise timing of these upregulations. Finally, the identification of the latter group of genes sheds light on potential cell cycle targets for modulating EMT trajectories. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
14. USP11 promotes prostate cancer progression by up-regulating AR and c-Myc activity.
- Author
-
Pornour, Majid, Hee-Young Jeon, Hyunju Ryu, Khadka, Sudeep, Rui Xu, Hegang Chen, Hussain, Arif, Hung-Ming Lam, Zhihao Zhuang, Htoo Zarni Oo, Gleave, Martin, Xuesen Dong, Qianben Wang, Barbieri, Christopher, and Jianfei Qi
- Subjects
CASTRATION-resistant prostate cancer ,ANDROGEN receptors ,GENE expression ,PROTEIN stability ,GENETIC transcription - Abstract
Androgen receptor (AR) is a main driver for castration-resistant prostate cancer (CRPC). c-Myc is an oncogene underlying prostate tumorigenesis. Here, we find that the deubiquitinase USP11 targets both AR and c-Myc in prostate cancer (PCa). USP11 expression was up-regulated in metastatic PCa and CRPC. USP11 knockdown (KD) significantly inhibited PCa cell growth. Our RNA-seq studies revealed AR and c-Myc as the top transcription factors altered after USP11 KD. ChIP-seq analysis showed that either USP11 KD or replacement of endogenous USP11 with a catalytic-inactive USP11 mutant significantly decreased chromatin binding by AR and c-Myc. We find that USP11 employs two mechanisms to up-regulate AR and c-Myc levels: namely, deubiquitination of AR and c-Myc proteins to increase their stability and deubiquitination of H2A-K119Ub, a repressive histone mark, on promoters of AR and c-Myc genes to increase their transcription. AR and c-Myc reexpression in USP11-KD PCa cells partly rescued cell growth defects. Thus, our studies reveal a tumor-promoting role for USP11 in aggressive PCa through upregulation of AR and c-Myc activities and support USP11 as a potential target against PCa. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
15. HLA-C expression in extravillous trophoblasts is determined by an ELF3-NLRP2/NLRP7 regulatory axis.
- Author
-
Bowen Gu, Gia-Han Le, Herrera, Sebastian, Blair, Steven J., Meissner, Torsten B., and Strominger, Jack L.
- Subjects
HLA histocompatibility antigens ,FETAL growth retardation ,GENE expression ,TRANSCRIPTION factors ,GENETIC regulation - Abstract
The distinct human leukocyte antigen (HLA) class I expression pattern of human extravillous trophoblasts (EVT) endows them with unique tolerogenic properties that enable successful pregnancy. Nevertheless, how this process is elaborately regulated remains elusive. Previously, E74 like ETS transcription factor 3 (ELF3) was identified to govern high-level HLA-C expression in EVT. In the present study, ELF3 is found to bind to the enhancer region of two adjacent NOD-like receptor (NLR) genes, NLR family pyrin domain-containing 2 and 7 (NLRP2, NLRP7). Notably, our analysis of ELF3-deficient JEG-3 cells, a human choriocarcinoma cell line widely used to study EVT biology, suggests that ELF3 transactivates NLRP7 while suppressing the expression of NLRP2. Moreover, we find that NLRP2 and NLRP7 have opposing effects on HLA-C expression, thus implicating them in immune evasion at the maternal-fetal interface. We confirmed that NLRP2 suppresses HLA-C levels and described a unique role for NLRP7 in promoting HLA-C expression in JEG-3. These results suggest that these two NLR genes, which arose via gene duplication in primates, are fine-tuned by ELF3 yet have acquired divergent functions to enable proper expression levels of HLA-C in EVT, presumably through modulating the degradation kinetics of IkBα. Targeting the ELF3-NLRP2/NLRP7-HLA-C axis may hold therapeutic potential for managing pregnancy-related disorders, such as recurrent hydatidiform moles and fetal growth restriction, and thus improve placental development and pregnancy outcomes. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
16. APC/C prevents a noncanonical order of cyclin/CDK activity to maintain CDK4/6 inhibitor-induced arrest.
- Author
-
Mouery, Brandon L., Baker, Eliyambuya M., Liu Mei, Wolff, Samuel C., Mills, Christine A., Fleifel, Dalia, Mulugeta, Nebyou, Herring, Laura E., and Cook, Jeanette Gowen
- Subjects
CYCLIN-dependent kinases ,DNA synthesis ,DNA replication ,GENE expression ,PROTEOLYSIS - Abstract
Regulated cell cycle progression ensures homeostasis and prevents cancer. In proliferating cells, premature S phase entry is avoided by the E3 ubiquitin ligase anaphasepromoting complex/cyclosome (APC/C), although the APC/C substrates whose degradation restrains G1 -S progression are not fully known. The APC/C is also active in arrested cells that exited the cell cycle, but it is not clear whether APC/C maintains all types of arrest. Here, by expressing the APC/C inhibitor, EMI1, we show that APC/C activity is essential to prevent S phase entry in cells arrested by pharmacological cyclin-dependent kinases 4 and 6 (CDK4/6) inhibition (Palbociclib). Thus, active protein degradation is required for arrest alongside repressed cell cycle gene expression. The mechanism of rapid and robust arrest bypass from inhibiting APC/C involves CDKs acting in an atypical order to inactivate retinoblastoma-mediated E2F repression. Inactivating APC/C first causes mitotic cyclin B accumulation which then promotes cyclin A expression. We propose that cyclin A is the key substrate for maintaining arrest because APC/C-resistant cyclin A, but not cyclin B, is sufficient to induce S phase entry. Cells bypassing arrest from CDK4/6 inhibition initiate DNA replication with severely reduced origin licensing. The simultaneous accumulation of S phase licensing inhibitors, such as cyclin A and geminin, with G1 licensing activators disrupts the normal order of G1-S progression. As a result, DNA synthesis and cell proliferation are profoundly impaired. Our findings predict that cancers with elevated EMI1 expression will tend to escape CDK4/6 inhibition into a premature, underlicensed S phase and suffer enhanced genome instability. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
17. Mitochondrial antioxidants abate SARS-COV-2 pathology in mice.
- Author
-
Guarnieri, Joseph W., Lie, Timothy, Soto Albrecht, Yentli E., Hewin, Peter, Jurado, Kellie A., Widjaja, Gabrielle A., Yi Zhu, McManus, Meagan J., Kilbaugh, Todd J., Keith, Kelsey, Potluri, Prasanth, Taylor, Deanne, Angelin, Alessia, Murdock, Deborah G., and Wallace, Douglas C.
- Subjects
SARS-CoV-2 ,ANGIOTENSIN converting enzyme ,MITOCHONDRIAL DNA ,VIRAL proteins ,GENE expression - Abstract
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection inhibits mito-chondrial oxidative phosphorylation (OXPHOS) and elevates mitochondrial reactive oxygen species (ROS, mROS) which activates hypoxia-inducible factor-1alpha (HIF-1a), shifting metabolism toward glycolysis to drive viral biogenesis but also causing the release of mitochondrial DNA (mtDNA) and activation of innate immunity. To determine whether mitochondrially targeted antioxidants could mitigate these viral effects, we challenged mice expressing human angiotensin-converting enzyme 2 (ACE2) with SARS-CoV-2 and intervened using transgenic and pharmacological mitochondrially targeted catalytic antioxidants. Transgenic expression of mitochondrially targeted catalase (mCAT) or systemic treatment with EUK8 decreased weight loss, clinical severity, and circulating levels of mtDNA; as well as reduced lung levels of HIF-1α, viral proteins, and inflammatory cytokines. RNA-sequencing of infected lungs revealed that mCAT and Eukarion 8 (EUK8) up-regulated OXPHOS gene expression and down-regulated HIF-1α and its target genes as well as innate immune gene expression. These data demonstrate that SARS-CoV-2 pathology can be mitigated by catalytically reducing mROS, potentially providing a unique host-directed pharmacological therapy for COVID-19 which is not subject to viral mutational resistance. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
18. Caenorhabditis elegans RIG-I-like receptor DRH-1 signals via CARDs to activate antiviral immunity in intestinal cells.
- Author
-
Batachari, Lakshmi E., Dai, Alyssa Y., and Troemel, Emily R.
- Subjects
RNA interference ,SMALL interfering RNA ,CAENORHABDITIS elegans ,DOUBLE-stranded RNA ,GENE expression - Abstract
Upon sensing viral RNA, mammalian RIG-I-like receptors (RLRs) activate downstream signals using caspase activation and recruitment domains (CARDs), which ultimately promote transcriptional immune responses that have been well studied. In contrast, the downstream signaling mechanisms for invertebrate RLRs are much less clear. For example, the Caenorhabditis elegans RLR DRH-1 lacks annotated CARDs and up-regulates the distinct output of RNA interference. Here, we found that similar to mammal RLRs, DRH-1 signals through two tandem CARDs (2CARD) to induce a transcriptional immune response. Expression of DRH-1(2CARD) alone in the intestine was sufficient to induce immune gene expression, increase viral resistance, and promote thermotolerance, a phenotype previously associated with immune activation in C. elegans. We also found that DRH-1 is required in the intestine to induce immune gene expression, and we demonstrate subcellular colocalization of DRH-1 puncta with double-stranded RNA inside the cytoplasm of intestinal cells upon viral infection. Altogether, our results reveal mechanistic and spatial insights into antiviral signaling in C. elegans, highlighting unexpected parallels in RLR signaling between C. elegans and mammals. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
19. The MKK3 MAPK cascade integrates temperature and after-ripening signals to modulate seed germination.
- Author
-
Masahiko Otani, Ryo Tojo, Regnard, Sarah, Lipeng Zheng, Takumi Hoshi, Suzuha Ohmori, Natsuki Tachibana, Tomohiro Sano, Shizuka Koshimizu, Kazuya Ichimura, Colcombet, Jean, and Naoto Kawakami
- Subjects
GERMINATION ,SEED quality ,LOCUS (Genetics) ,MITOGEN-activated protein kinases ,GENE expression ,ABSCISIC acid - Abstract
The timing of seed germination is controlled by the combination of internal dormancy and external factors. Temperature is a major environmental factor for seed germination. The permissive temperature range for germination is narrow in dormant seeds and expands during after-ripening (AR) (dormancy release). Quantitative trait loci analyses of preharvest sprouting in cereals have revealed that MKK3, a mitogen-activated protein kinase (MAPK) cascade protein, is a negative regulator of grain dormancy. Here, we show that the MAPKKK19/20-MKK3-MPK1/2/7/14 cascade modulates the germination temperature range in Arabidopsis seeds by elevating the germinability of the seeds at sub-and supraoptimal temperatures. The expression of MAPKKK19 and MAPKKK20 is induced around optimal temperature for germination in after-ripened seeds but repressed in dormant seeds. MPK7 activation depends on the expression levels of MAPKKK19/20, with expression occurring under conditions permissive for germination. Abscisic acid (ABA) and gibberellin (GA) are two major phytohormones which are involved in germination control. Activation of the MKK3 cascade represses ABA biosynthesis enzyme gene expression and induces expression of ABA catabolic enzyme and GA biosynthesis enzyme genes, resulting in expansion of the germinable temperature range. Our data demonstrate that the MKK3 cascade integrates temperature and AR signals to phytohormone metabolism and seed germination. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
20. Membrane association of active genes organizes the chloroplast nucleoid structure.
- Author
-
Palomar, V. Miguel, Yoonjin Cho, Sho Fujii, Rothi, M. Hafiz, Jaksich, Sarah, Ji-Hee Min, Schlachter, Adriana N., Joyful Wang, Zhengde Liu, and Wierzbicki, Andrzej T.
- Subjects
CHLOROPLAST DNA ,GENETIC transcription ,ORGANELLE formation ,GENE expression ,GENES ,SPIDER silk - Abstract
DNA is organized into chromatin-like structures that support the maintenance and regulation of genomes. A unique and poorly understood form of DNA organization exists in chloroplasts, which are organelles of endosymbiotic origin responsible for photosynthesis. Chloroplast genomes, together with associated proteins, form membrane-less structures known as nucleoids. The internal arrangement of the nucleoid, molecular mechanisms of DNA organization, and connections between nucleoid structure and gene expression remain mostly unknown. We show that Arabidopsis thaliana chloroplast nucleoids have a unique sequence-specific organization driven by DNA binding to the thylakoid membranes. DNA associated with the membranes has high protein occupancy, has reduced DNA accessibility, and is highly transcribed. In contrast, genes with low levels of transcription are further away from the membranes, have lower protein occupancy, and have higher DNA accessibility. Membrane association of active genes relies on the pattern of transcription and proper chloroplast development. We propose a speculative model that transcription organizes the chloroplast nucleoid into a transcriptionally active membrane-associated core and a less active periphery. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
21. The DNA damage response of Escherichia coli, revisited: Differential gene expression after replication inhibition.
- Author
-
Sass, Thalia H. and Lovett, Susan T.
- Subjects
DNA repair ,GENE expression ,ESCHERICHIA coli ,TRANSCRIPTION factors ,IRON in the body - Abstract
In 1967, in this journal, Evelyn Witkin proposed the existence of a coordinated DNA damage response in Escherichia coli, which later came to be called the "SOS response." We revisited this response using the replication inhibitor azidothymidine (AZT) and RNA-Seq analysis and identified several features. We confirm the induction of classic Save our ship (SOS) loci and identify several genes, including many of the pyrimidine pathway, that have not been previously demonstrated to be DNA damage-inducible. Despite a strong dependence on LexA, these genes lack LexA boxes and their regulation by LexA is likely to be indirect via unknown factors. We show that the transcription factor "stringent starvation protein" SspA is as important as LexA in the regulation of AZT-induced genes and that the genes activated by SspA change dramatically after AZT exposure. Our experiments identify additional LexA-independent DNA damage inducible genes, including 22 small RNA genes, some of which appear to activated by SspA. Motility and chemotaxis genes are strongly down-regulated by AZT, possibly as a result of one of more of the small RNAs or other transcription factors such as AppY and GadE, whose expression is elevated by AZT. Genes controlling the iron siderophore, enterobactin, and iron homeostasis are also strongly induced, independent of LexA. We confirm that IraD antiadaptor protein is induced independent of LexA and that a second antiadaptor, IraM is likewise strongly AZT-inducible, independent of LexA, suggesting that RpoS stabilization via these antiadaptor proteins is an integral part of replication stress tolerance. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
22. Genomic structural variation contributes to evolved changes in gene expression in high-altitude Tibetan sheep.
- Author
-
Xiaolong Liang, Qijiao Duan, Bowen Li, Yinjia Wang, Yueting Bu, Yonglu Zhang, Zhuoran Kuang, Leyan Mao, Xuan An, Huihua Wang, Xiaojie Yang, Na Wan, Zhilong Feng, Wei Shen, Weilan Miao, Jiaqi Chen, Sanyuan Liu, Storz, Jay F., Jianquan Liu, and Nevo, Eviatar
- Subjects
GENE expression ,SHEEP breeds ,SHEEP ,CHROMOSOME inversions ,GENETIC models - Abstract
Tibetan sheep were introduced to the Qinghai Tibet plateau roughly 3,000 B.P., making this species a good model for investigating genetic mechanisms of high-altitude adaptation over a relatively short timescale. Here, we characterize genomic structural variants (SVs) that distinguish Tibetan sheep from closely related, low-altitude Hu sheep, and we examine associated changes in tissue-specific gene expression. We document differentiation between the two sheep breeds in frequencies of SVs associated with genes involved in cardiac function and circulation. In Tibetan sheep, we identified high-frequency SVs in a total of 462 genes, including EPAS1, PAPSS2, and PTPRD. Single-cell RNA-Seq data and luciferase reporter assays revealed that the SVs had cis-acting effects on the expression levels of these three genes in specific tissues and cell types. In Tibetan sheep, we identified a high-frequency chromosomal inversion that exhibited modified chromatin architectures relative to the noninverted allele that predominates in Hu sheep. The inversion harbors several genes with altered expression patterns related to heart protection, brown adipocyte proliferation, angiogenesis, and DNA repair. These findings indicate that SVs represent an important source of genetic variation in gene expression and may have contributed to high-altitude adaptation in Tibetan sheep. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
23. NF-κB activation is a turn on for vaccinia virus phosphoprotein A49 to turn off NF-κB activation.
- Author
-
Neidel, Sarah, Ren, Hongwei, Torres, Alice A., and Smith, Geoffrey L.
- Subjects
VACCINIA ,PROTEINS ,CELL proliferation ,GENE expression ,PHOSPHORYLATION - Abstract
Vaccinia virus protein A49 inhibits NF-κB activation by molecular mimicry and has a motif near the N terminus that is conserved in IκBa, ß-catenin, HIV Vpu, and some other proteins. This motif contains two serines, and for IκBa and ß-catenin, phosphorylation of these serines enables recognition by the E3 ubiquitin ligase ß-TrCP. Binding of IκBa and ß-catenin by ß-TrCP causes their ubiquitylation and thereafter proteasome-mediated degradation. In contrast, HIV Vpu and VACV A49 are not degraded. This paper shows that A49 is phosphorylated at serine 7 but not serine 12 and that this is necessary and sufficient for binding ß-TrCP and antagonism of NF-κB. Phosphorylation of A49 S7 occurs when NF-κB signaling is activated by addition of IL-1ß or overexpression of TRAF6 or IKKß, the kinase needed for IκBa phosphorylation. Thus, A49 shows beautiful biological regulation, for it becomes an NF-κB antagonist upon activation of NF-κB signaling. The virulence of viruses expressing mutant A49 proteins or lacking A49 (vΔA49) was tested. vΔA49 was attenuated compared with WT, but viruses expressing A49 that cannot bind ß-TrCP or bind ß-TrCP constitutively had intermediate virulence. So A49 promotes virulence by inhibiting NF-κB activation and by another mechanism independent of S7 phosphorylation and NF-κB antagonism. Last, a virus lacking A49 was more immunogenic than the WT virus. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
24. SI Correction.
- Subjects
CELL adhesion molecules ,GREEN fluorescent protein ,GENE expression ,GENETIC vectors ,MOLECULAR size - Abstract
This document is a correction to the supporting information for a scientific article on inhibitory synapse development. The correction addresses an image that was included twice in the supporting information. The article describes experiments and assays conducted to study the interaction between proteins involved in synapse formation. The results were analyzed using various techniques and statistically analyzed for significance. The supplementary information includes figures and tables related to the experiments, providing useful data for researchers studying synaptic development and function. [Extracted from the article]
- Published
- 2024
- Full Text
- View/download PDF
25. Deciphering the functional specialization of whole-brain spatiomolecular gradients in the adult brain.
- Author
-
Vogel, Jacob W., Alexander-Bloch, Aaron F., Wagstyl, Konrad, Bertolero, Maxwell A., Markello, Ross D., Pines, Adam, Sydnor, Valerie J., Diaz-Papkovich, Alex, Hansen, Justine Y., Evans, Alan C., Bernhardt, Boris, Misic, Bratislav, Satterthwaite, Theodore D., and Seidlitz, Jakob
- Subjects
RELIEF models ,GENE expression ,ADULTS ,TRANSCRIPTION factors ,NEURAL development - Abstract
Cortical arealization arises during neurodevelopment from the confluence of molecular gradients representing patterned expression of morphogens and transcription factors. However, whether similar gradients are maintained in the adult brain remains unknown. Here, we uncover three axes of topographic variation in gene expression in the adult human brain that specifically capture previously identified rostral-caudal, dorsal-ventral, and medial-lateral axes of early developmental patterning. The interaction of these spatiomolecular gradients i) accurately reconstructs the position of brain tissue samples, ii) delineates known functional territories, and iii) can model the topographical variation of diverse cortical features. The spatiomolecular gradients are distinct from canonical cortical axes differentiating the primary sensory cortex from the association cortex, but radiate in parallel with the axes traversed by local field potentials along the cortex. We replicate all three molecular gradients in three independent human datasets as well as two nonhuman primate datasets and find that each gradient shows a distinct developmental trajectory across the lifespan. The gradients are composed of several well-known transcription factors (e.g., PAX6 and SIX3), and a small set of genes shared across gradients are strongly enriched for multiple diseases. Together, these results provide insight into the developmental sculpting of functionally distinct brain regions, governed by three robust transcriptomic axes embedded within brain parenchyma. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
26. The role of mitochondria in sex-and age-specific gene expression in a species without sex chromosomes.
- Author
-
Ning Li, Flanagan, Ben A., and Edmands, Suzanne
- Subjects
SEX chromosomes ,GENE expression ,MITOCHONDRIAL DNA ,MITOCHONDRIA ,RNA sequencing ,ANIMAL industry ,TRANSACTIONAL sex - Abstract
Mitochondria perform an array of functions, many of which involve interactions with gene products encoded by the nucleus. These mitochondrial functions, particularly those involving energy production, can be expected to differ between sexes and across ages. Here, we measured mitochondrial effects on sex-and age-specific gene expression in parental and reciprocal F1 hybrids between allopatric populations of Tigriopus californicus with over 20% mitochondrial DNA divergence. Because the species lacks sex chromosomes, sex-biased mitochondrial effects are not confounded by the effects of sex chromosomes. Results revealed pervasive sex differences in mitochondrial effects, including effects on energetics and aging involving nuclear interactions throughout the genome. Using single-individual RNA sequencing, sex differences were found to explain more than 80% of the variance in gene expression. Males had higher expression of mitochondrial genes and mitochondrially targeted proteins (MTPs) involved in oxidative phosphorylation (OXPHOS), while females had elevated expression of non-OXPHOS MTPs, indicating strongly sex-dimorphic energy metabolism at the whole organism level. Comparison of reciprocal F1 hybrids allowed insights into the nature of mito-nuclear interactions, showing both mitochondrial effects on nuclear expression, and nuclear effects on mitochondrial expression. While based on a small set of crosses, sex-specific increases in mitochondrial expression with age were associated with longer life. Network analyses identified nuclear components of strong mito-nuclear interactions and found them to be sexually dimorphic. These results highlight the profound impact of mitochondria and mito-nuclear interactions on sex-and age-specific gene expression. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
27. How total mRNA influences cell growth.
- Author
-
Calabrese, Ludovico, Ciandrini, Luca, and Cosentino Lagomarsino, Marco
- Subjects
CELL growth ,MESSENGER RNA ,RNA polymerases ,GENE expression ,PROTEIN expression - Abstract
Experimental observations tracing back to the 1960s imply that ribosome quantities play a prominent role in determining a cell's growth. Nevertheless, in biologically relevant scenarios, growth can also be influenced by the levels of mRNA and RNA polymerase. Here, we construct a quantitative model of biosynthesis providing testable scenarios for these situations. The model explores a theoretically motivated regime where RNA polymerases compete for genes and ribosomes for transcripts and gives general expressions relating growth rate, mRNA concentrations, ribosome, and RNA polymerase levels. On general grounds, the model predicts how the fraction of ribosomes in the proteome depends on total mRNA concentration and inspects an underexplored regime in which the trade-off between transcript levels and ribosome abundances sets the cellular growth rate. In particular, we show that the model predicts and clarifies three important experimental observations, in budding yeast and Escherichia coli bacteria: i) that the growth-rate cost of unneeded protein expression can be affected by mRNA levels, ii) that resource optimization leads to decreasing trends in mRNA levels at slow growth, and iii) that ribosome allocation may increase, stay constant, or decrease, in response to transcription-inhibiting antibiotics. Since the data indicate that a regime of joint limitation may apply in physiological conditions and not only to perturbations, we speculate that this regime is likely self-imposed. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
28. Clocking out and letting go to unleash green biotech applications in a photosynthetic host.
- Author
-
Yao Xu, Jabbur, Maria Luísa, Tetsuya Mori, Young, Jamey D., and Johnson, Carl Hirschie
- Subjects
GENE expression ,PHOTOSYNTHETIC bacteria ,ALTERNATIVE fuels ,GENETIC transcription ,ESCHERICHIA coli - Abstract
Cyanobacteria are photosynthetic bacteria whose gene expression patterns are globally regulated by their circadian (daily) clocks. Due to their ability to use sunlight as their energy source, they are also attractive hosts for "green" production of pharmaceuticals, renewable fuels, and chemicals. However, despite the application of traditional genetic tools such as the identification of strong promoters to enhance the expression of heterologous genes, cyanobacteria have lagged behind other microorganisms such as Escherichia coli and yeast as economically efficient cell factories. The previous approaches have ignored large-scale constraints within cyanobacterial metabolic networks on transcription, predominantly the pervasive control of gene expression by the circadian (daily) clock. Here, we show that reprogramming gene expression by releasing circadian repressor elements in the transcriptional regulatory pathways coupled with inactivation of the central oscillating mechanism enables a dramatic enhancement of expression in cyanobacteria of heterologous genes encoding both catalytically active enzymes and polypeptides of biomedical significance. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
29. DNA lesion bypass and the stochastic dynamics of transcription-coupled repair.
- Author
-
Nicholson, Michael D., Anderson, Craig J., Odom, Duncan T., Aitken, Sarah J., and Taylor, Martin S.
- Subjects
EXCISION repair ,DNA damage ,RNA polymerase II ,GENE expression ,RNA polymerases - Abstract
DNA base damage is a major source of oncogenic mutations and disruption to gene expression. The stalling of RNA polymerase II (RNAP) at sites of DNA damage and the subsequent triggering of repair processes have major roles in shaping the genome-wide distribution of mutations, clearing barriers to transcription, and minimizing the production of miscoded gene products. Despite its importance for genetic integrity, key mechanistic features of this transcription-coupled repair (TCR) process are controversial or unknown. Here, we exploited a well-powered in vivo mammalian model system to explore the mechanistic properties and parameters of TCR for alkylation damage at fine spatial resolution and with discrimination of the damaged DNA strand. For rigorous interpretation, a generalizable mathematical model of DNA damage and TCR was developed. Fitting experimental data to the model and simulation revealed that RNA polymerases frequently bypass lesions without triggering repair, indicating that small alkylation adducts are unlikely to be an efficient barrier to gene expression. Following a burst of damage, the efficiency of transcription-coupled repair gradually decays through gene bodies with implications for the occurrence and accurate inference of driver mutations in cancer. The reinitation of transcription from the repair site is not a general feature of transcription-coupled repair, and the observed data is consistent with reinitiation never taking place. Collectively, these results reveal how the directional but stochastic activity of TCR shapes the distribution of mutations following DNA damage. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
30. A conserved molecular logic for neurogenesis to gliogenesis switch in the cerebral cortex.
- Author
-
Xiaoyi G. Liang, Hoang, Kendy, Meyerink, Brandon L., Kc, Pratiksha, Paraiso, Kitt, Li Wang, Jones, Ian R., Yue Zhang, Katzman, Sol, Finn, Thomas S., Tsyporin, Jeremiah, Fangyuan Qu, Zhaoxu Chen, Visel, Axel, Kriegstein, Arnold, Yin Shen, Pilaz, Louis-Jan, and Bin Chen
- Subjects
CEREBRAL cortex ,NEURAL stem cells ,GENE expression ,NEUROGENESIS ,NEUROGLIA - Abstract
During development, neural stem cells in the cerebral cortex, also known as radial glial cells (RGCs), generate excitatory neurons, followed by production of cortical macroglia and inhibitory neurons that migrate to the olfactory bulb (OB). Understanding the mechanisms for this lineage switch is fundamental for unraveling how proper numbers of diverse neuronal and glial cell types are controlled. We and others recently showed that Sonic Hedgehog (Shh) signaling promotes the cortical RGC lineage switch to generate cortical oligodendrocytes and OB interneurons. During this process, cortical RGCs generate intermediate progenitor cells that express critical gliogenesis genes Ascll, Egfr, and Olig2. The increased Ascll expression and appearance of Egfr
+ and Olig2+ cortical progenitors are concurrent with the switch from excitatory neurogenesis to gliogenesis and OB interneuron neurogenesis in the cortex. While Shh signaling promotes Olig2 expression in the developing spinal cord, the exact mechanism for this transcriptional regulation is not known. Furthermore, the transcriptional regulation of Olig2 and Egfr has not been explored. Here, we show that in cortical progenitor cells, multiple regu-latory programs, including Pax6 and Gli3, prevent precocious expression of Olig2, a gene essential for production of cortical oligodendrocytes and astrocytes. We identify multiple enhancers that control Olig2 expression in cortical progenitors and show that the mechanisms for regulating Olig2 expression are conserved between the mouse and human. Our study reveals evolutionarily conserved regulatory logic controlling the lineage switch of cortical neural stem cells. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
31. Species-wide quantitative transcriptomes and proteomes reveal distinct genetic control of gene expression variation in yeast.
- Author
-
Teyssonnière, Elie Marcel, Trébulle, Pauline, Muenzner, Julia, Loegler, Victor, Ludwig, Daniela, Amari, Fatma, Mülleder, Michael, Friedrich, Anne, Jing Hou, Ralser, Markus, and Schacherer, Joseph
- Subjects
GENE expression ,TRANSCRIPTOMES ,GENETIC variation ,YEAST ,PROTEOMICS - Abstract
Gene expression varies between individuals and corresponds to a key step linking genotypes to phenotypes. However, our knowledge regarding the species-wide genetic control of protein abundance, including its dependency on transcript levels, is very limited. Here, we have determined quantitative proteomes of a large population of 942 diverse natural Saccharomyces cerevisiae yeast isolates. We found that mRNA and protein abundances are weakly correlated at the population gene level. While the protein coexpression network recapitulates major biological functions, differential expression patterns reveal proteomic signatures related to specific populations. Comprehensive genetic association analyses highlight that genetic variants associated with variation in protein (pQTL) and transcript (eQTL) levels poorly overlap (3%). Our results demonstrate that transcriptome and proteome are governed by distinct genetic bases, likely explained by protein turnover. It also highlights the importance of integrating these different levels of gene expression to better understand the genotype-phenotype relationship. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
32. Liver cancer development driven by the AP- 1/c- Jun-Fra- 2 dimer through c- Myc.
- Author
-
Bakiri, Latifa, Hasenfuss, Sebastian C., Guío-Carrión, Ana, Thomsen, Martin K., and Wagner, Peter Hasselblatt Erwin F.
- Subjects
LIVER cancer ,CARCINOGENESIS ,TRANSGENE expression ,TRANSCRIPTION factors ,GENE expression - Abstract
Hepatocellular carcinoma (HCC) is a leading cause of cancer- related death. HCC incidence is on the rise, while treatment options remain limited. Thus, a better understanding of the molecular pathways involved in HCC development has become a priority to guide future therapies. While previous studies implicated the Activator Protein- 1 (AP- 1) (Fos/Jun) transcription factor family members c- Fos and c- Jun in HCC formation, the contribution of Fos- related antigens (Fra- ) 1 and 2 is unknown. Here, we show that hepatocyte- restricted expression of a single chain c- Jun-Fra- 2 protein, which functionally mimics the c- Jun/Fra- 2 AP- 1 dimer, results in spontaneous HCC formation in c- Jun-Fra- 2hep mice. Several hallmarks of human HCC, such as cell cycle dysregulation and the expression of HCC markers are observed in liver tumors arising in c- Jun-Fra- 2hep mice. Tumorigenesis occurs in the context of mild inflammation, low- grade fibrosis, and Pparγ- driven dyslipidemia. ubsequent analyses revealed increased expression of c- Myc, evidently under direct regulation by AP- 1 through a conserved distal 3 enhancer. Importantly, c- Jun-Fra- 2- induced tumors revert upon switching off transgene expression, suggesting oncogene addiction to the c- Jun-Fra- 2 transgene. Tumors escaping reversion maintained c- Myc and c- Myc target gene expression, likely due to increased c- Fos. Interfering with c- Myc in established tumors using the Bromodomain and Extra- Terminal motif inhibitor JQ- 1 diminished liver tumor growth in c- Jun-Fra- 2 mutant mice. Thus, our data establish c- Jun-Fra- 2hep mice as a model to study liver tumorigenesis and identify the c- Jun/Fra- 2- Myc interaction as a potential target to improve HCC patient stratification and/or therapy. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
33. Organ- delimited gene regulatory networks provide high accuracy in candidate transcription factor selection across diverse processes.
- Author
-
Ranjan, Rajeev, Srijan, Sonali, Balekuttira, Somaiah, Agarwal, Tina, Ramey, Melissa, Dobbins, Madison, Kuhn, Rachel, Wang, Xiaojin, HudsonYing Li, Karen, and Varala, Kranthi
- Subjects
GENE regulatory networks ,TRANSCRIPTION factors ,ACID phosphatase ,LIPID synthesis ,GENE expression - Abstract
Organ- specific gene expression datasets that include hundreds to thousands of experiments allow the reconstruction of organ- level gene regulatory networks (GRNs). However, creating such datasets is greatly hampered by the requirements of extensive and tedious manual curation. Here, we trained a supervised classification model that can accurately classify the organ- of- origin for a plant transcriptome. This K- Nearest Neighbor- based multiclass classifier was used to create organ- specific gene expression datasets for the leaf, root, shoot, flower, and seed in Arabidopsis thaliana. A GRN inference approach was used to determine the: i. influential transcription factors (TFs) in each organ and, ii. most influential TFs for specific biological processes in that organ. These genome- wide, organ- delimited GRNs (OD- GRNs), recalled many known regulators of organ development and processes operating in those organs. Importantly, many previously unknown TF regulators were uncovered as potential regulators of these processes. As a proof- of- concept, we focused on experimentally validating the predicted TF regulators of lipid biosynthesis in seeds, an important food and biofuel trait. Of the top 20 predicted TFs, eight are known regulators of seed oil content, e.g., WRI1, LEC1, FUS3. Importantly, we validated our prediction of MybS2, TGA4, SPL12, AGL18, and DiV2 as regulators of seed lipid biosynthesis. We elucidated the molecular mechanism of MybS2 and show that it induces purple acid phosphatase family genes and lipid synthesis genes to enhance seed lipid content. This general approach has the potential to be extended to any species with sufficiently large gene expression datasets to find unique regulators of any trait- of- interest. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
34. A mammalian tripartite enhancer cluster controls hypothalamic Pomc expression, food intake, and body weight.
- Author
-
Rojo, Daniela, Hael, Clara E., Soria, Agustina, de Souza, Flávio S. J., Low, Malcolm J., Franchin, Lucía F., and Rubinstein, Marcelo
- Subjects
GENE expression ,FOOD consumption ,BODY weight ,MOLECULAR evolution ,BINDING sites - Abstract
Food intake and energy balance are tightly regulated by a group of hypothalamic arcuate neurons expressing the proopiomelanocortin (POMC) gene. In mammals, arcuate- specific POMC expression is driven by two cis- acting transcriptional enhancers known as nPE1 and nPE2. Because mutant mice lacking these two enhancers still showed hypothalamic Pomc mRNA, we searched for additional elements contributing to arcuate Pomc expression. By combining molecular evolution with reporter gene expression in transgenic zebrafish and mice, here, we identified a mammalian arcuate- specific Pomc enhancer that we named nPE3, carrying several binding sites also present in nPE1 and nPE2 for transcription factors known to activate neuronal Pomc expression, such as ISL1, NKX2.1, and ERα. We found that nPE3 originated in the lineage leading to placental mammals and remained under purifying selection in all mammalian orders, although it was lost in Simiiformes (monkeys, apes, and humans) following a unique segmental deletion event. Interestingly, ablation of nPE3 from the mouse genome led to a drastic reduction (>70%) in hypothalamic Pomc mRNA during development and only moderate (<33%) in adult mice. Comparison between double (nPE1 and nPE2) and triple (nPE1, nPE2, and nPE3) enhancer mutants revealed the relative contribution of nPE3 to hypothalamic Pomc expression and its importance in the control of food intake and adiposity in male and female mice. Altogether, these results demonstrate that nPE3 integrates a tripartite cluster of partially redundant enhancers that originated upon a triple convergent evolutionary process in mammals and that is critical for hypothalamic Pomc expression and body weight homeostasis. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
35. The scaffolding protein AKAP12 regulates mRNA localization and translation.
- Author
-
Smith, Madeleine R., Naeli, Parisa, Jafarnejad, Seyed M., and Costa, Guilherme
- Subjects
SCAFFOLD proteins ,GENETIC translation ,MESSENGER RNA ,PROTEIN kinases ,GENE expression - Abstract
Regulation of subcellular messenger (m)RNA localization is a fundamental biological mechanism, which adds a spatial dimension to the diverse layers of post- transcriptional control of gene expression. The cellular compartment in which mRNAs are located may define distinct aspects of the encoded proteins, ranging from production rate and complex formation to localized activity. Despite the detailed roles of localized mRNAs that have emerged over the past decades, the identity of factors anchoring mRNAs to subcellular domains remains ill- defined. Here, we used an unbiased method to profile the RNA- bound proteome in migrating endothelial cells (ECs) and discovered that the plasma membrane (PM)--associated scaffolding protein A- kinase anchor protein (AKAP)12 interacts with various mRNAs, including transcripts encoding kinases with Actin remodeling activity. In particular, AKAP12 targets a transcript coding for the kinase Abelson Tyrosine- Protein Kinase 2 (ABL2), which we found to be necessary for adequate filopodia formation and angiogenic sprouting. Moreover, we demonstrate that AKAP12 is necessary for anchoring ABL2 mRNA to the PM and show that in the absence of AKAP12, the translation efficiency of ABL2 mRNA is reduced. Altogether, our work identified a unique post- transcriptional function for AKAP12 and sheds light into mechanisms of spatial control of gene expression. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
36. Structural rearrangements in the nucleus localize latent HIV proviruses to a perinucleolar compartment supportive of reactivation.
- Author
-
Kizito, Fredrick, Nguyen, Kien, Mbonye, Uri, Shukla, Meenakshi, Luttge, Benjamin, Checkley, Mary Ann, Agaponova, Anna, Leskov, Konstantin, and Karn, Jonathan
- Subjects
HIV-positive persons ,PROMOTERS ,HIV ,GENE expression ,NUCLEAR pore complex - Abstract
This document provides a comprehensive overview of a study on the localization and transcription of HIV proviruses in T cells. The study used various techniques, including Cas-FISH imaging and CRISPR-Cas9 technology, to investigate the subnuclear distribution of HIV DNA and RNA, as well as the structural rearrangements in the nucleus that occur during the transition into quiescence and reactivation. The findings suggest that the perinucleolar compartment (PNC) plays a crucial role in HIV transcription and reactivation, and provide valuable insights into the molecular mechanisms of HIV latency. The document also includes detailed protocols and references for further research. [Extracted from the article]
- Published
- 2024
- Full Text
- View/download PDF
37. The Myc-associated zinc finger protein epigenetically controls expression of interferon-γ-stimulated genes by recruiting STAT1 to chromatin.
- Author
-
Tiaojiang Xiao, Xin Li, and Felsenfeld, Gary
- Subjects
ZINC-finger proteins ,GENE expression ,STAT proteins ,CHROMATIN ,GENETIC regulation ,NATURAL products ,PROSTATE cancer patients - Abstract
The MYC-Associated Zinc Finger Protein (MAZ) plays important roles in chromatin organization and gene transcription regulation. Dysregulated expression of MAZ causes diseases, such as glioblastoma, breast cancer, prostate cancer, and liposarcoma. Previously, it has been reported that MAZ controls the proinflammatory response in colitis and colon cancer via STAT3 signaling, suggesting that MAZ is involved in regulating immunity-related pathways. However, the molecular mechanism underlying this regulation remains elusive. Here, we investigate the regulatory effect of MAZ on interferon-gamma (IFN-1)- stimulated genes via STAT1, a protein that plays an essential role in immune responses to viral, fungal, and mycobacterial pathogens. We demonstrate that about 80% of occupied STAT1-binding sites colocalize with occupied MAZ-binding sites in HAP1/K562 cells after IFN-1 stimulation. MAZ depletion significantly reduces STAT1 binding in the genome. By analyzing genome-wide gene expression profiles in the RNA-Seq data, we show that MAZ depletion significantly suppresses a subset of the immune response genes, which include the IFN-stimulated genes IRF8 and Absent in Melanoma 2. Furthermore, we find that MAZ controls expression of the immunity-related genes by changing the epigenetic landscape in chromatin. Our study reveals an important role for MAZ in regulating immune-related gene expression. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
38. Hox gene-specific cellular targeting using split intein Trojan exons.
- Author
-
Diao, Fengqiu, Vasudevan, Deeptha, Heckscher, Ellie S., and White, Benjamin H.
- Subjects
TRANSCRIPTION factors ,MODULAR design ,GENE expression ,NEURAL circuitry ,MUTAGENS ,DROSOPHILA - Abstract
The Trojan exon method, which makes use of intronically inserted T2A-Gal4 cassettes, has been widely used in Drosophila to create thousands of gene-specific Gal4 driver lines. These dual-purpose lines provide genetic access to specific cell types based on their expression of a native gene while simultaneously mutating one allele of the gene to enable loss-of-function analysis in homozygous animals. While this dual use is often an advantage, the truncation mutations produced by Trojan exons are sometimes deleterious in heterozygotes, perhaps by creating translation products with dominant negative effects. Such mutagenic effects can cause developmental lethality as has been observed with genes encoding essential transcription factors. Given the importance of transcription factors in specifying cell type, alternative techniques for generating specific Gal4 lines that target them are required. Here, we introduce a modified Trojan exon method that retains the targeting fidelity and plug-and-play modularity of the original method but mitigates its mutagenic effects by exploiting the self-splicing capabilities of split inteins. "Split Intein Trojan exons" (siTrojans) ensure that the two truncation products generated from the interrupted allele of the native gene are trans-spliced to create a full-length native protein. We demonstrate the efficacy of siTrojans by generating a comprehensive toolkit of Gal4 and Split Gal4 lines for the segmentally expressed Hox transcription factors and illustrate their use in neural circuit mapping by targeting neurons according to their position along the anterior-posterior axis. Both the method and the Hox gene-specific toolkit introduced here should be broadly useful. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
39. Reversal of C9orf72 mutation-induced transcriptional dysregulation and pathology in cultured human neurons by allele-specific excision.
- Author
-
Sachdev, Aradhana, Gill, Kamaljot, Sckaff, Maria, Birk, Alisha M., Arogundade, Olubankole Aladesuyi, Brown, Katherine A., Chouhan, Runvir S., Issagholian-Lewin, Patrick Oliver, Patel, Esha, Watry, Hannah L., Bernardi, Mylinh T., Keough, Kathleen C., Yu-Chih Tsai, Smith, Alec Simon Tulloch, Conklin, Bruce R., and Clelland, Claire Dudley
- Subjects
GENE expression ,INDUCED pluripotent stem cells ,DEMENTIA patients ,SINGLE nucleotide polymorphisms ,GENETIC regulation ,AMYOTROPHIC lateral sclerosis - Abstract
Efforts to genetically reverse C9orf72 pathology have been hampered by our incomplete understanding of the regulation of this complex locus. We generated five different genomic excisions at the C9orf72 locus in a patient-derived induced pluripotent stem cell (iPSC) line and a non-diseased wild-type (WT) line (11 total isogenic lines), and examined gene expression and pathological hallmarks of C9 frontotemporal dementia/amyotrophic lateral sclerosis in motor neurons differentiated from these lines. Comparing the excisions in these isogenic series removed the confounding effects of different genomic backgrounds and allowed us to probe the effects of specific genomic changes. A coding single nucleotide polymorphism in the patient cell line allowed us to distinguish transcripts from the normal vs. mutant allele. Using digital droplet PCR (ddPCR), we determined that transcription from the mutant allele is upregulated at least 10-fold, and that sense transcription is independently regulated from each allele. Surprisingly, excision of the WT allele increased pathologic dipeptide repeat poly-GP expression from the mutant allele. Importantly, a single allele was sufficient to supply a normal amount of protein, suggesting that the C9orf72 gene is haplo-sufficient in induced motor neurons. Excision of the mutant repeat expansion reverted all pathology (RNA abnormalities, dipeptide repeat production, and TDP-43 pathology) and improved electrophysiological function, whereas silencing sense expression did not eliminate all dipeptide repeat proteins, presumably because of the antisense expression. These data increase our understanding of C9orf72 gene regulation and inform gene therapy approaches, including antisense oligonucleotides (ASOs) and CRISPR gene editing. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
40. Posttranscriptional regulation of FAN1 by miR-124-3p at rs3512 underlies onset-delaying genetic modification in Huntington's disease.
- Author
-
Kyung-Hee Kim, Eun Pyo Hong, Yukyeong Lee, McLean, Zachariah L., Elezi, Emanuela, Lee, Ramee, Seung Kwak, McAllister, Branduff, Massey, Thomas H., Lobanov, Sergey, Holmans, Peter, Orth, Michael, Ciosi, Marc, Monckton, Darren G., Long, Jeffrey D., Lucente, Diane, Wheeler, Vanessa C., MacDonald, Marcy E., Gusella, James F., and Jong-Min Lee
- Subjects
HUNTINGTON disease ,LOCUS (Genetics) ,GENE expression ,GENETIC variation ,MESSENGER RNA - Abstract
Many Mendelian disorders, such as Huntington's disease (HD) and spinocerebellar ataxias, arise from expansions of CAG trinucleotide repeats. Despite the clear genetic causes, additional genetic factors may influence the rate of those monogenic disorders. Notably, genome-wide association studies discovered somewhat expected modifiers, particularly mismatch repair genes involved in the CAG repeat instability, impacting age at onset of HD. Strikingly, FAN1, previously unrelated to repeat instability, produced the strongest HD modification signals. Diverse FAN1 haplotypes independently modify HD, with rare genetic variants diminishing DNA binding or nuclease activity of the FAN1 protein, hastening HD onset. However, the mechanism behind the frequent and the most significant onset-delaying FAN1 haplotype lacking missense variations has remained elusive. Here, we illustrated that a microRNA acting on 3'-UTR (untranslated region) SNP rs3512, rather than transcriptional regulation, is responsible for the significant FAN1 expression quantitative trait loci signal and allelic imbalance in FAN1 messenger ribonucleic acid (mRNA), accounting for the most significant and frequent onset-delaying modifier haplotype in HD. Specifically, miR-124-3p selectively targets the reference allele at rs3512, diminishing the stability of FAN1 mRNA harboring that allele and consequently reducing its levels. Subsequent validation analyses, including the use of antagomir and 3'-UTR reporter vectors with swapped alleles, confirmed the specificity of miR-124-3p at rs3512. Together, these findings indicate that the alternative allele at rs3512 renders the FAN1 mRNA less susceptible to miR-124-3p-mediated posttranscriptional regulation, resulting in increased FAN1 levels and a subsequent delay in HD onset by mitigating CAG repeat instability. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
41. Developmental trajectories of GABAergic cortical interneurons are sequentially modulated by dynamic FoxG1 expression levels.
- Author
-
Goichi Miyoshi, Yoshifumi Ueta, Yuki Yagasaki, Yusuke Kishi, Fishell, Gord, Machold, Robert P., and Mariko Miyata
- Subjects
GENE expression ,INTERNEURONS ,AUTISM spectrum disorders - Abstract
GABAergic inhibitory interneurons, originating from the embryonic ventral forebrain territories, traverse a convoluted migratory path to reach the neocortex. These interneuron precursors undergo sequential phases of tangential and radial migration before settling into specific laminae during differentiation. Here, we show that the developmental trajectory of FoxG1 expression is dynamically controlled in these interneuron precursors at critical junctures of migration. By utilizing mouse genetic strategies, we elucidate the pivotal role of precise changes in FoxG1 expression levels during interneuron specification and migration. Our findings underscore the gene dosage-dependent function of FoxG1, aligning with clinical observations of FOXG1 haploinsufficiency and duplication in syndromic forms of autism spectrum disorders. In conclusion, our results reveal the finely tuned developmental clock governing cortical interneuron development, driven by temporal dynamics and the dose-dependent actions of FoxG1. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
42. The genetic regulatory architecture and epigenomic basis for age-related changes in rattlesnake venom.
- Author
-
Hogan, Michael P., Holding, Matthew L., Nystrom, Gunnar S., Colston, Timothy J., Bartlett, Daniel A., Mason, Andrew J., Ellsworth, Schyler A., Rautsaw, Rhett M., Lawrence, Kylie C., Strickland, Jason L., Bing He, Fraser, Peter, Margres, Mark J., Gilbert, David M., Gibbs, H. Lisle, Parkinson, Christopher L., and Rokyta, Darin R.
- Subjects
VENOM ,BIOLOGICAL systems ,GENE expression ,RATTLESNAKES ,LIFE history theory - Abstract
Developmental phenotypic changes can evolve under selection imposed by age- and size-related ecological differences. Many of these changes occur through programmed alterations to gene expression patterns, but the molecular mechanisms and generegulatory networks underlying these adaptive changes remain poorly understood. Many venomous snakes, including the eastern diamondback rattlesnake (Crotalus adamanteus), undergo correlated changes in diet and venom expression as snakes grow larger with age, providing models for identifying mechanisms of timed expression changes that underlie adaptive life history traits. By combining a highly contiguous, chromosome-level genome assembly with measures of expression, chromatin accessibility, and histone modifications, we identified cis-regulatory elements and transregulatory factors controlling venom ontogeny in the venom glands of C. adamanteus. Ontogenetic expression changes were significantly correlated with epigenomic changes within genes, immediately adjacent to genes (e.g., promoters), and more distant from genes (e.g., enhancers). We identified 37 candidate transcription factors (TFs), with the vast majority being up-regulated in adults. The ontogenetic change is largely driven by an increase in the expression of TFs associated with growth signaling, transcriptional activation, and circadian rhythm/biological timing systems in adults with corresponding epigenomic changes near the differentially expressed venom genes. However, both expression activation and repression contributed to the composition of both adult and juvenile venoms, demonstrating the complexity and potential evolvability of gene regulation for this trait. Overall, given that age-based trait variation is common across the tree of life, we provide a framework for understanding generegulatory-network-driven life-history evolution more broadly. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
43. Transcriptional elongation control of hypoxic response.
- Author
-
Soliman, Shimaa Hassan AbdelAziz, Iwanaszko, Marta, Bin Zheng, Gold, Sarah, Howard, Benjamin Charles, Das, Madhurima, Chakrabarty, Ram Prosad, Chandel, Navdeep S., and Shilatifard, Ali
- Subjects
RNA polymerase II ,GENETIC regulation - Abstract
The release of paused RNA polymerase II (RNAPII) from promoter-proximal regions is tightly controlled to ensure proper regulation of gene expression. The elongation factor PTEF-b is known to release paused RNAPII via phosphorylation of the RNAPII C-terminal domain by its cyclin-dependent kinase component, CDK9. However, the signal and stress-specific roles of the various RNAPII-associated macromolecular complexes containing PTEF-b/CDK9 are not yet clear. Here, we identify and characterize the CDK9 complex required for transcriptional response to hypoxia. Contrary to previous reports, our data indicate that a CDK9 complex containing BRD4 but not AFF1/4 is essential for this hypoxic stress response. We demonstrate that BRD4 bromodomains (BET) are dispensable for the release of paused RNAPII at hypoxia-activated genes and that BET inhibition by JQ1 is insufficient to impair hypoxic gene response. Mechanistically, we demonstrate that the C-terminal region of BRD4 is required for Polymerase-Associated Factor-1 Complex (PAF1C) recruitment to establish an elongation-competent RNAPII complex at hypoxia-responsive genes. PAF1C disruption using a small-molecule inhibitor (iPAF1C) impairs hypoxia-induced, BRD4-mediated RNAPII release. Together, our results provide insight into potentially targetable mechanisms that control the hypoxia-responsive transcriptional elongation. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
44. Timeless noncoding DNA contains cell-type preferential enhancers important for proper Drosophila circadian regulation.
- Author
-
Dingbang Ma, Ojha, Pranav, Yu, Albert D., Araujo, Maisa S., Weifei Luo, Keefer, Evelyn, Díaz, Madelen M., Meilin Wu, Joiner, William J., Abruzzi, Katharine C., and Rosbash, Michael
- Subjects
NON-coding DNA ,GENE expression ,MOLECULAR clock ,CLOCK genes ,DROSOPHILA ,NATURAL products - Abstract
To address the contribution of transcriptional regulation to Drosophila clock gene expression and to behavior, we generated a series of CRISPR-mediated deletions within two regions of the circadian gene timeless (tim), an intronic E-box region and an upstream E-box region that are both recognized by the key transcription factor Clock (Clk) and its heterodimeric partner Cycle. The upstream deletions but not an intronic deletion dramatically impact tim expression in fly heads; the biggest upstream deletion reduces peak RNA levels and tim RNA cycling amplitude to about 15% of normal, and there are similar effects on tim protein (TIM). The cycling amplitude of other clock genes is also strongly reduced, in these cases due to increases in trough levels. These data underscore the important contribution of the upstream E-box enhancer region to tim expression and of TIM to clock gene transcriptional repression in fly heads. Surprisingly, tim expression in clock neurons is only modestly affected by the biggest upstream deletion and is similarly affected by a deletion of the intronic E-box region. This distinction between clock neurons and glia is paralleled by a dramatically enhanced accessibility of the intronic enhancer region within clock neurons. This distinctive feature of tim chromatin was revealed by ATAC-seq (assay for transposase-accessible chromatin with sequencing) assays of purified neurons and glia as well as of fly heads. The enhanced cell type-specific accessibility of the intronic enhancer region explains the resilience of clock neuron tim expression and circadian behavior to deletion of the otherwise more prominent upstream tim E-box region. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
45. Biclustering with heterogeneous variance.
- Author
-
Guanhua Chen, Sullivan, Patrick F., and Kosorok, Michael R.
- Subjects
CANCER ,GENETICS ,ANALYSIS of variance ,GENE expression ,HEURISTIC - Abstract
In cancer research, as in all of medicine, it is important to classify patients into etiologically and therapeutically relevant subtypes to improve diagnosis and treatment. One way to do this is to use clustering methods to find subgroups of homogeneous individuals based on genetic profiles together with heuristic clinical analysis. A notable drawback of existing clustering methods is that they ignore the possibility that the variance of gene expression profile measurements can be heterogeneous across subgroups, and methods that do not consider heterogeneity of variance can lead to inaccurate subgroup prediction. Research has shown that hyper-variability is a common feature among cancer subtypes. In this paper, we present a statistical approach that can capture both mean and variance structure in genetic data. We demonstrate the strength of our method in both synthetic data and in two cancer data sets. In particular, our method confirms the hypervariability of methylation level in cancer patients, and it detects clearer subgroup patterns in lung cancer data. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
46. Disentangling the pseudoknots of toxin translation.
- Author
-
Jagodnik, Jonathan, Darfeuille, Fabien, and Guillier, Maude
- Subjects
BIOLOGICAL evolution ,NUCLEIC acids ,ESCHERICHIA coli toxins ,GENE expression ,ANTISENSE RNA - Abstract
The article explores the translation of toxin genes in bacteria's toxin-antitoxin systems (TA). These systems consist of a toxic protein and its antitoxin antidote. The study focuses on the timP-TimR T1TA system in Salmonella and reveals that a pseudoknot structure in the timP mRNA activates translation. The antitoxin TimR destabilizes the pseudoknot structure, inhibiting translation. The findings shed light on the role of RNA dynamics in gene expression and offer insights into alternative ways toxin mRNA can be activated. Understanding the control of toxin and antitoxin synthesis can provide valuable information about their physiological functions. [Extracted from the article]
- Published
- 2024
- Full Text
- View/download PDF
47. IL7 increases targeted lipid nanoparticle-mediated mRNA expression in T cells in vitro and in vivo by enhancing T cell protein translation.
- Author
-
Tilsed, Caitlin M., Sadiq, Barzan A., Papp, Tyler E., Areesawangkit, Phurin, Kenji Kimura, Noguera-Orteg, Estela, Scholler, John, Cerda, Nicholas, Aghajanian, Haig, Bot, Adrian, Mui, Barbara, Ying Tam, Weissman, Drew, June, Carl H., Albelda, Steven M., and Parhiz, Hamideh
- Subjects
T cells ,GENE expression ,INTRAVENOUS injections ,PROTEIN expression ,LIPIDS - Abstract
The use of lipid nanoparticles (LNP) to encapsulate and deliver mRNA has become an important therapeutic advance. In addition to vaccines, LNP-mRNA can be used in many other applications. For example, targeting the LNP with anti-CD5 antibodies (CD5/tLNP) can allow for efficient delivery of mRNA payloads to T cells to express protein. As the percentage of protein expressing T cells induced by an intravenous injection of CD5/tLNP is relatively low (4-20%), our goal was to find ways to increase mRNA-induced translation efficiency. We showed that T cell activation using an anti-CD3 antibody improved protein expression after CD5/tLNP transfection in vitro but not in vivo. T cell health and activation can be increased with cytokines, therefore, using mCherry mRNA as a reporter, we found that culturing either mouse or human T cells with the cytokine IL7 significantly improved protein expression of delivered mRNA in both CD4
+ and CD8+ T cells in vitro. By pre-treating mice with systemic IL7 followed by tLNP administration, we observed significantly increased mCherry protein expression by T cells in vivo. Transcriptomic analysis of mouse T cells treated with IL7 in vitro revealed enhanced genomic pathways associated with protein translation. Improved translational ability was demonstrated by showing increased levels of protein expression after electroporation with mCherry mRNA in T cells cultured in the presence of IL7, but not with IL2 or IL15. These data show that IL7 selectively increases protein translation in T cells, and this property can be used to improve expression of tLNP-delivered mRNA in vivo. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
48. Heterogeneous elasticity drives ripening and controls bursting kinetics of transcriptional condensates.
- Author
-
Lingyu Meng, Sheng Mao, and Jie Lin
- Subjects
LOGNORMAL distribution ,GENE expression ,OSTWALD ripening ,ELASTICITY ,GENETIC regulation - Abstract
Many biomolecular condensates, including transcriptional condensates, are formed in elastic mediums. In this work, we study the nonequilibrium condensate dynamics in a chromatin-like environment modeled as a heterogeneous elastic medium. We demonstrate that the ripening process in such an elastic medium exhibits a temporal power-law scaling of the average condensate radius, depending on the local stiffness distribution and different from Ostwald ripening. Moreover, we incorporate an active process to model the dissolution of transcriptional condensates upon RNA accumulation. Intriguingly, three types of kinetics of condensate growth emerge, corresponding to constitutively expressed, transcriptional-bursting, and silenced genes. Furthermore, the simulated burst frequency decreases exponentially with the local stiffness, through which we infer a lognormal distribution of local stiffness in living cells using the transcriptome-wide distribution of burst frequency. Under the inferred stiffness distribution, the simulated distributions of bursting kinetic parameters agree reasonably well with the experimental data. Our findings reveal the interplay between biomolecular condensates and elastic mediums, yielding far-reaching implications for gene expression. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
49. Aging and comprehensive molecular profiling in acute myeloid leukemia.
- Author
-
Jian-Feng Li, Wen-Yan Cheng, Xiang-Jie Lin, Li-Jun Wen, Kai Wang, Yong-Mei Zhu, Hong-Ming Zhu, Xin-Jie Chen, Yu-Liang Zhang, Wei Yin, Jia-Nan Zhang, Xiao Yi, Fan Zhang, Xiang-Qin Weng, Sheng-Yue Wang, Lu Jiang, Hui-Yi Wu, Jia-Qi Ren, Xiao-Jing Lin, and Niu Qiao
- Subjects
ACUTE myeloid leukemia ,HEMATOLOGIC malignancies ,MYELODYSPLASTIC syndromes ,GENE fusion ,GENE expression - Abstract
Acute myeloid leukemia (AML) is an aging-related and heterogeneous hematopoietic malignancy. In this study, a total of 1,474 newly diagnosed AML patients with RNA sequencing data were enrolled, and targeted or whole exome sequencing data were obtained in 94% cases. The correlation of aging-related factors including age and clonal hematopoiesis (CH), gender, and genomic/transcriptomic profiles (gene fusions, genetic mutations, and gene expression networks or pathways) was systematically analyzed. Overall, AML patients aged 60 y and older showed an apparently dismal prognosis. Alongside age, the frequency of gene fusions defined in the World Health Organization classification decreased, while the positive rate of gene mutations, especially CH-related ones, increased. Additionally, the number of genetic mutations was higher in gene fusion--negative (GF-) patients than those with GF. Based on the status of CH- and myelodysplastic syndromes (MDS)-related mutations, three mutant subgroups were identified among the GF-AML cohort, namely, CH-AML, CH-MDS-AML, and other GF-AML. Notably, CH-MDS-AML demonstrated a predominance of elderly and male cases, cytopenia, and significantly adverse clinical outcomes. Besides, gene expression networks including HOXA/B, platelet factors, and inflammatory responses were most striking features associated with aging and poor prognosis in AML. Our work has thus unraveled the intricate regulatory circuitry of interactions among different age, gender, and molecular groups of AML. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
50. CHD7 and SOX2 act in a common gene regulatory network during mammalian semicircular canal and cochlear development.
- Author
-
Jingxia Gao, Skidmore, Jennifer M., Cimerman, Jelka, Ritter, K. Elaine, Jingyun Qiu, Wilson, Lindsey M. Q., Raphael, Yehoash, Kwan, Kelvin Y., and Martin, Donna M.
- Subjects
GENE regulatory networks ,FORKHEAD transcription factors ,SEMICIRCULAR canals ,INNER ear ,VESTIBULAR apparatus ,GENE expression - Abstract
Inner ear morphogenesis requires tightly regulated epigenetic and transcriptional control of gene expression. CHD7, an ATP-dependent chromodomain helicase DNA-binding protein, and SOX2, an SRY-related HMG box pioneer transcription factor, are known to contribute to vestibular and auditory system development, but their genetic interactions in the ear have not been explored. Here, we analyzed inner ear development and the transcriptional regulatory landscapes in mice with variable dosages of Chd7 and/or Sox2. We show that combined haploinsufficiency for Chd7 and Sox2 results in reduced otic cell proliferation, severe malformations of semicircular canals, and shortened cochleae with ectopic hair cells. Examination of mice with conditional, inducible Chd7 loss by Sox2
CreER reveals a critical period (~E9.5) of susceptibility in the inner ear to combined Chd7 and Sox2 loss. Data from genome-wide RNA-sequencing and CUT&Tag studies in the otocyst show that CHD7 regulates Sox2 expression and acts early in a gene regulatory network to control expression of key otic patterning genes, including Pax2 and Otx2. CHD7 and SOX2 directly bind independently and cooperatively at transcription start sites and enhancers to regulate otic progenitor cell gene expression. Together, our findings reveal essential roles for Chd7 and Sox2 in early inner ear development and may be applicable for syndromic and other forms of hearing or balance disorders. [ABSTRACT FROM AUTHOR]- Published
- 2024
- Full Text
- View/download PDF
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.