79 results on '"Wittkopp PJ"'
Search Results
2. Cis-regulatory elements: molecular mechanisms and evolutionary processes underlying divergence.
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Wittkopp PJ, Kalay G, Wittkopp, Patricia J, and Kalay, Gizem
- Abstract
Cis-regulatory sequences, such as enhancers and promoters, control development and physiology by regulating gene expression. Mutations that affect the function of these sequences contribute to phenotypic diversity within and between species. With many case studies implicating divergent cis-regulatory activity in phenotypic evolution, researchers have recently begun to elucidate the genetic and molecular mechanisms that are responsible for cis-regulatory divergence. Approaches include detailed functional analysis of individual cis-regulatory elements and comparing mechanisms of gene regulation among species using the latest genomic tools. Despite the limited number of mechanistic studies published to date, this work shows how cis-regulatory activity can diverge and how studies of cis-regulatory divergence can address long-standing questions about the genetic mechanisms of phenotypic evolution. [ABSTRACT FROM AUTHOR]
- Published
- 2012
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3. A micro-RNA is the effector gene of a classic evolutionary hotspot locus.
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Tian S, Asano Y, Banerjee TD, Wee JLQ, Lamb A, Wang Y, Murugesan SN, Ui-Tei K, Wittkopp PJ, and Monteiro A
- Abstract
In Lepidoptera (butterflies and moths), the genomic region around the gene cortex is a 'hotspot' locus, repeatedly used to generate intraspecific melanic wing color polymorphisms across 100-million-years of evolution. However, the identity of the effector gene regulating melanic wing color within this locus remains unknown. Here, we show that none of the four candidate protein-coding genes within this locus, including cortex , serve as major effectors. Instead, a micro-RNA (miRNA), mir-193 , serves as the major effector across three deeply diverged lineages of butterflies, and its function is conserved in Drosophila . In Lepidoptera, mir-193 is derived from a gigantic long non-coding RNA, ivory , and it functions by directly repressing multiple pigmentation genes. We show that a miRNA can drive repeated instances of adaptive evolution in animals., Competing Interests: Competing interests: Authors declare that they have no competing interests.
- Published
- 2024
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4. Plasticity and environment-specific relationships between gene expression and fitness in Saccharomyces cerevisiae .
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Siddiq MA, Duveau F, and Wittkopp PJ
- Abstract
Phenotypic evolution is shaped by interactions between organisms and their environments. The environment influences how an organism's genotype determines its phenotype and how this phenotype affects its fitness. To better understand this dual role of the environment in the production and selection of phenotypic variation, we empirically determined and compared the genotype-phenotype-fitness relationship for mutant strains of the budding yeast Saccharomyces cerevisiae in four environments. Specifically, we measured how mutations in the promoter of the metabolic gene TDH3 modified its expression level and affected its growth on media with four different carbon sources. In each environment, we observed a clear relationship between TDH3 expression level and fitness, but this relationship differed among environments. Genetic variants with similar effects on TDH3 expression in different environments often had different effects on fitness and vice versa. Such environment-specific relationships between phenotype and fitness can shape the evolution of phenotypic plasticity. The set of mutants we examined also allowed us to compare the effects of mutations disrupting binding sites for key transcriptional regulators and the TATA box, which is part of the core promoter sequence. Mutations disrupting the binding sites for the transcription factors had more variable effects on expression among environments than mutations disrupting the TATA box, yet mutations with the most environmentally variable effects on fitness were located in the TATA box. This observation suggests that mutations affecting different molecular mechanisms are likely to contribute unequally to regulatory sequence evolution in changing environments., Competing Interests: Competing Interest Statement: The authors have no competing interests to disclose.
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- 2024
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5. Divergence of Grainy head affects chromatin accessibility, gene expression, and embryonic viability in Drosophila melanogaster .
- Author
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Ertl HA, Bayala EX, Siddiq MA, and Wittkopp PJ
- Abstract
Pioneer factors are critical for gene regulation and development because they bind chromatin and make DNA more accessible for binding by other transcription factors. The pioneer factor Grainy head (Grh) is present across metazoans and has been shown to retain a role in epithelium development in fruit flies, nematodes, and mice despite extensive divergence in both amino acid sequence and length. Here, we investigate the evolution of Grh function by comparing the effects of the fly ( Drosophila melanogaster ) and worm ( Caenorhabditis elegans ) Grh orthologs on chromatin accessibility, gene expression, embryonic development, and viability in transgenic D. melanogaster . We found that the Caenorhabditis elegans ortholog rescued cuticle development but not full embryonic viability in Drosophila melanogaster grh null mutants. At the molecular level, the C. elegans ortholog only partially rescued chromatin accessibility and gene expression. Divergence in the disordered N-terminus of the Grh protein contributes to these differences in embryonic viability and molecular phenotypes. These data show how pioneer factors can diverge in sequence and function at the molecular level while retaining conserved developmental functions at the organismal level., Competing Interests: COMPETING INTERESTS No competing interests declared.
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- 2024
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6. Active compensation for changes in TDH3 expression mediated by direct regulators of TDH3 in Saccharomyces cerevisiae.
- Author
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Vande Zande P, Siddiq MA, Hodgins-Davis A, Kim L, and Wittkopp PJ
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- Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins genetics, Saccharomyces cerevisiae Proteins metabolism
- Abstract
Genetic networks are surprisingly robust to perturbations caused by new mutations. This robustness is conferred in part by compensation for loss of a gene's activity by genes with overlapping functions, such as paralogs. Compensation occurs passively when the normal activity of one paralog can compensate for the loss of the other, or actively when a change in one paralog's expression, localization, or activity is required to compensate for loss of the other. The mechanisms of active compensation remain poorly understood in most cases. Here we investigate active compensation for the loss or reduction in expression of the Saccharomyces cerevisiae gene TDH3 by its paralog TDH2. TDH2 is upregulated in a dose-dependent manner in response to reductions in TDH3 by a mechanism requiring the shared transcriptional regulators Gcr1p and Rap1p. TDH1, a second and more distantly related paralog of TDH3, has diverged in its regulation and is upregulated by another mechanism. Other glycolytic genes regulated by Rap1p and Gcr1p show changes in expression similar to TDH2, suggesting that the active compensation by TDH3 paralogs is part of a broader homeostatic response mediated by shared transcriptional regulators., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Vande Zande et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2023
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7. Interdisciplinary approaches to predicting evolutionary biology.
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Crocker J, Payne JL, Walczak AM, and Wittkopp PJ
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- Biological Evolution, Biology
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- 2023
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8. Contributions of mutation and selection to regulatory variation: lessons from the Saccharomyces cerevisiae TDH3 gene.
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Wittkopp PJ
- Subjects
- Mutation, Selection, Genetic, Polymorphism, Genetic, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins genetics, Saccharomyces cerevisiae Proteins metabolism
- Abstract
Heritable variation in gene expression is common within and among species and contributes to phenotypic diversity. Mutations affecting either cis - or trans -regulatory sequences controlling gene expression give rise to variation in gene expression, and natural selection acting on this variation causes some regulatory variants to persist in a population for longer than others. To understand how mutation and selection interact to produce the patterns of regulatory variation we see within and among species, my colleagues and I have been systematically determining the effects of new mutations on expression of the TDH3 gene in Saccharomyces cerevisiae and comparing them to the effects of polymorphisms segregating within this species. We have also investigated the molecular mechanisms by which regulatory variants act. Over the past decade, this work has revealed properties of cis - and trans -regulatory mutations including their relative frequency, effects, dominance, pleiotropy and fitness consequences. Comparing these mutational effects to the effects of polymorphisms in natural populations, we have inferred selection acting on expression level, expression noise and phenotypic plasticity. Here, I summarize this body of work and synthesize its findings to make inferences not readily discernible from the individual studies alone. This article is part of the theme issue 'Interdisciplinary approaches to predicting evolutionary biology'.
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- 2023
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9. Emerging questions in transcriptional regulation.
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Nora EP, Aerts S, Wittkopp PJ, Bussemaker HJ, Bulyk M, Sinha S, Zeitlinger J, Crocker J, and Fuxman Bass JI
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- Gene Expression Regulation genetics
- Abstract
What new questions can we ask about transcriptional regulation given recent developments in large-scale approaches?, Competing Interests: Declaration of interests J.Z. is an investigator at the Stowers Institute for Medical Research and also a professor (affiliate track) at the Department of Pathology and Laboratory Medicine at the University of Kansas Medical Center. H.J.B. is a co-founder and shareholder of Metric Biotechnologies, Inc. Columbia University has filed a patent on a technology tangentially related, on which H.J.B. is one of the inventors, to the topic of the Voices piece that H.J.B. contributed., (Copyright © 2023. Published by Elsevier Inc.)
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- 2023
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10. Active compensation for changes in TDH3 expression mediated by direct regulators of TDH3 in Saccharomyces cerevisiae .
- Author
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Zande PV and Wittkopp PJ
- Abstract
Genetic networks are surprisingly robust to perturbations caused by new mutations. This robustness is conferred in part by compensation for loss of a gene's activity by genes with overlapping functions, such as paralogs. Compensation occurs passively when the normal activity of one paralog can compensate for the loss of the other, or actively when a change in one paralog's expression, localization, or activity is required to compensate for loss of the other. The mechanisms of active compensation remain poorly understood in most cases. Here we investigate active compensation for the loss or reduction in expression of the Saccharomyces cerevisiae gene TDH3 by its paralogs TDH1 and TDH2. TDH1 and TDH2 are upregulated in a dose-dependent manner in response to reductions in TDH3 by a mechanism requiring the shared transcriptional regulators Gcr1p and Rap1p. Other glycolytic genes regulated by Rap1p and Gcr1p show changes in expression similar to TDH2 , suggesting that the active compensation by TDH3 paralogs is part of a broader homeostatic response mediated by shared transcriptional regulators.
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- 2023
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11. Differential Grainy head binding correlates with variation in chromatin structure and gene expression in Drosophila melanogaster.
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Ertl HA, Hill MS, and Wittkopp PJ
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- Animals, DNA-Binding Proteins genetics, Transcription Factors genetics, Transcription Factors metabolism, Chromatin genetics, Chromatin metabolism, Gene Expression, Drosophila melanogaster genetics, Drosophila melanogaster metabolism, Drosophila Proteins genetics, Drosophila Proteins metabolism
- Abstract
Phenotypic evolution is often caused by variation in gene expression resulting from altered gene regulatory mechanisms. Genetic variation affecting chromatin remodeling has been identified as a potential source of variable gene expression; however, the roles of specific chromatin remodeling factors remain unclear. Here, we address this knowledge gap by examining the relationship between variation in gene expression, variation in chromatin structure, and variation in binding of the pioneer factor Grainy head between imaginal wing discs of two divergent strains of Drosophila melanogaster and their F
1 hybrid. We find that (1) variation in Grainy head binding is mostly due to sequence changes that act in cis but are located outside of the canonical Grainy head binding motif, (2) variation in Grainy head binding correlates with changes in chromatin accessibility, and (3) this variation in chromatin accessibility, coupled with variation in Grainy head binding, correlates with variation in gene expression in some cases but not others. Interactions among these three molecular layers is complex, but these results suggest that genetic variation affecting the binding of pioneer factors contributes to variation in chromatin remodeling and the evolution of gene expression., (© 2022. The Author(s).)- Published
- 2022
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12. Network Topology Can Explain Differences in Pleiotropy Between Cis- and Trans-regulatory Mutations.
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Vande Zande P and Wittkopp PJ
- Subjects
- Mutation, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, Phenotype, Evolution, Molecular, Saccharomyces cerevisiae Proteins genetics, Saccharomyces cerevisiae Proteins metabolism
- Abstract
A mutation's degree of pleiotropy (i.e., the number of traits it alters) is predicted to impact the probability of the mutation being detrimental to fitness. For mutations that impact gene expression, mutations acting in cis have been hypothesized to generally be less pleiotropic than mutations affecting the same gene's expression in trans, suggesting that cis-regulatory mutations should be less deleterious and more likely to fix over evolutionary time. Here, we use expression and fitness data from Saccharomyces cerevisiae gene deletion strains to test these hypotheses. By treating deletion of each gene as a cis-regulatory mutation affecting its own expression and deletions of other genes affecting expression of this focal gene as trans-regulatory mutations, we find that cis-acting mutations do indeed tend to be less pleiotropic than trans-acting mutations affecting expression of the same gene. This pattern was observed for the vast majority of genes in the data set and could be explained by the topology of the regulatory network controlling gene expression. Comparing the fitness of cis- and trans-acting mutations affecting expression of the same gene also confirmed that trans-acting deletions tend to be more deleterious. These findings provide strong support for pleiotropy playing a role in the preferential fixation of cis-regulatory alleles over evolutionary time., (© The Author(s) 2022. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution.)
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- 2022
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13. Mechanisms of regulatory evolution in yeast.
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Siddiq MA and Wittkopp PJ
- Subjects
- Genome, Saccharomyces cerevisiae genetics, Evolution, Molecular
- Abstract
Studies of regulatory variation in yeast - at the level of new mutations, polymorphisms within a species, and divergence between species - have provided great insight into the molecular and evolutionary processes responsible for the evolution of gene expression in eukaryotes. The increasing ease with which yeast genomes can be manipulated and expression quantified in a high-throughput manner has recently accelerated mechanistic studies of cis- and trans-regulatory variation at multiple evolutionary timescales. These studies have, for example, identified differences in the properties of cis- and trans-acting mutations that affect their evolutionary fate, experimentally characterized the molecular mechanisms through which cis- and trans-regulatory variants act, and illustrated how regulatory networks can diverge between species with or without changes in gene expression., (Copyright © 2022 Elsevier Ltd. All rights reserved.)
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- 2022
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14. Pleiotropic effects of trans-regulatory mutations on fitness and gene expression.
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Vande Zande P, Hill MS, and Wittkopp PJ
- Subjects
- Alleles, Gene Expression, Mutation, Evolution, Molecular, Gene Expression Regulation, Fungal, Genetic Fitness, Genetic Variation, Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) genetics, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins genetics
- Abstract
Variation in gene expression arises from cis- and trans-regulatory mutations, which contribute differentially to expression divergence. We compare the impacts on gene expression and fitness resulting from cis- and trans-regulatory mutations in Saccharomyces cerevisiae , with a focus on the TDH3 gene. We use the effects of cis-regulatory mutations to infer effects of trans-regulatory mutations attributable to impacts beyond the focal gene, revealing a distribution of pleiotropic effects. Cis- and trans-regulatory mutations had different effects on gene expression with pleiotropic effects of trans-regulatory mutants affecting expression of genes both in parallel to and downstream of the focal gene. The more widespread and deleterious effects of trans-regulatory mutations we observed are consistent with their decreasing relative contribution to expression differences over evolutionary time.
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- 2022
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15. Distinct genetic architectures underlie divergent thorax, leg, and wing pigmentation between Drosophila elegans and D. gunungcola.
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Massey JH, Li J, Stern DL, and Wittkopp PJ
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- Alleles, Animals, Male, Pigmentation genetics, Species Specificity, Thorax, Drosophila genetics, Drosophila Proteins genetics
- Abstract
Pigmentation divergence between Drosophila species has emerged as a model trait for studying the genetic basis of phenotypic evolution, with genetic changes contributing to pigmentation differences often mapping to genes in the pigment synthesis pathway and their regulators. These studies of Drosophila pigmentation have tended to focus on pigmentation changes in one body part for a particular pair of species, but changes in pigmentation are often observed in multiple body parts between the same pair of species. The similarities and differences of genetic changes responsible for divergent pigmentation in different body parts of the same species thus remain largely unknown. Here we compare the genetic basis of pigmentation divergence between Drosophila elegans and D. gunungcola in the wing, legs, and thorax. Prior work has shown that regions of the genome containing the pigmentation genes yellow and ebony influence the size of divergent male-specific wing spots between these two species. We find that these same two regions of the genome underlie differences in leg and thorax pigmentation; however, divergent alleles in these regions show differences in allelic dominance and epistasis among the three body parts. These complex patterns of inheritance can be explained by a model of evolution involving tissue-specific changes in the expression of Yellow and Ebony between D. elegans and D. gunungcola., (© 2021. The Author(s), under exclusive licence to The Genetics Society.)
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- 2021
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16. Mutational sources of trans -regulatory variation affecting gene expression in Saccharomyces cerevisiae .
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Duveau F, Vande Zande P, Metzger BP, Diaz CJ, Walker EA, Tryban S, Siddiq MA, Yang B, and Wittkopp PJ
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- Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins metabolism, Gene Expression Regulation, Fungal, Mutation, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins genetics
- Abstract
Heritable variation in a gene's expression arises from mutations impacting cis - and trans -acting components of its regulatory network. Here, we investigate how trans -regulatory mutations are distributed within the genome and within a gene regulatory network by identifying and characterizing 69 mutations with trans -regulatory effects on expression of the same focal gene in Saccharomyces cerevisiae . Relative to 1766 mutations without effects on expression of this focal gene, we found that these trans -regulatory mutations were enriched in coding sequences of transcription factors previously predicted to regulate expression of the focal gene. However, over 90% of the trans -regulatory mutations identified mapped to other types of genes involved in diverse biological processes including chromatin state, metabolism, and signal transduction. These data show how genetic changes in diverse types of genes can impact a gene's expression in trans , revealing properties of trans -regulatory mutations that provide the raw material for trans -regulatory variation segregating within natural populations., Competing Interests: FD, PV, BM, CD, EW, ST, MS, BY No competing interests declared, PW Senior editor, eLife, (© 2021, Duveau et al.)
- Published
- 2021
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17. Molecular and evolutionary processes generating variation in gene expression.
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Hill MS, Vande Zande P, and Wittkopp PJ
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- Alleles, Gene Regulatory Networks genetics, Genetic Variation genetics, Mutation genetics, Evolution, Molecular, Gene Expression Regulation genetics, Selection, Genetic genetics
- Abstract
Heritable variation in gene expression is common within and between species. This variation arises from mutations that alter the form or function of molecular gene regulatory networks that are then filtered by natural selection. High-throughput methods for introducing mutations and characterizing their cis- and trans-regulatory effects on gene expression (particularly, transcription) are revealing how different molecular mechanisms generate regulatory variation, and studies comparing these mutational effects with variation seen in the wild are teasing apart the role of neutral and non-neutral evolutionary processes. This integration of molecular and evolutionary biology allows us to understand how the variation in gene expression we see today came to be and to predict how it is most likely to evolve in the future.
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- 2021
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18. Genetic architecture of a body colour cline in Drosophila americana.
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Sramkoski LL, McLaughlin WN, Cooley AM, Yuan DC, John A, and Wittkopp PJ
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- Animals, Color, North America, Pigmentation genetics, Drosophila genetics, Drosophila Proteins
- Abstract
Phenotypic variation within a species is often structured geographically in clines. In Drosophila americana, a longitudinal cline for body colour exists within North America that appears to be due to local adaptation. The tan and ebony genes have been hypothesized to contribute to this cline, with alleles of both genes that lighten body colour found in D. americana. These alleles are similar in sequence and function to the allele fixed in D. americana's more lightly pigmented sister species, Drosophila novamexicana. Here, we examine the frequency and geographic distribution of these D. novamexicana-like alleles in D. americana. Among alleles from over 100 strains of D. americana isolated from 21 geographic locations, we failed to identify additional alleles of tan or ebony with as much sequence similarity to D. novamexicana as the D. novamexicana-like alleles previously described. However, using genetic analysis of 51 D. americana strains derived from 20 geographic locations, we identified one new allele of ebony and one new allele of tan segregating in D. americana that are functionally equivalent to the D. novamexicana allele. An additional 5 alleles of tan also showed marginal evidence of functional similarity. Given the rarity of these alleles, however, we conclude that they are unlikely to be driving the pigmentation cline. Indeed, phenotypic distributions of the 51 backcross populations analysed indicate a more complex genetic architecture, with diversity in the number and effects of loci altering pigmentation observed both within and among populations of D. americana. This genetic heterogeneity poses a challenge to association studies and genomic scans for clinal variation, but might be common in natural populations., (© 2020 John Wiley & Sons Ltd.)
- Published
- 2020
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19. ebony affects pigmentation divergence and cuticular hydrocarbons in Drosophila americana and D. novamexicana .
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Lamb AM, Wang Z, Simmer P, Chung H, and Wittkopp PJ
- Abstract
Drosophila pigmentation has been a fruitful model system for understanding the genetic and developmental mechanisms underlying phenotypic evolution. For example, prior work has shown that divergence of the tan gene contributes to pigmentation differences between two members of the virilis group: Drosophila novamexicana , which has a light yellow body color, and D. americana , which has a dark brown body color. Quantitative trait locus (QTL) mapping and expression analysis has suggested that divergence of the ebony gene might also contribute to pigmentation differences between these two species. Here, we directly test this hypothesis by using CRISPR/Cas9 genome editing to generate ebony null mutants in D. americana and D. novamexicana and then using reciprocal hemizygosity testing to compare the effects of each species' ebony allele on pigmentation. We find that divergence of ebony does indeed contribute to the pigmentation divergence between species, with effects on both the overall body color as well as a difference in pigmentation along the dorsal abdominal midline. Motivated by recent work in D. melanogaster , we also used the ebony null mutants to test for effects of ebony on cuticular hydrocarbon (CHC) profiles. We found that ebony affects CHC abundance in both species, but does not contribute to qualitative differences in the CHC profiles between these two species. Additional transgenic resources for working with D. americana and D. novamexicana, such as white mutants of both species and yellow mutants in D. novamexicana , were generated in the course of this work and are also described. Taken together, this study advances our understanding of loci contributing to phenotypic divergence and illustrates how the latest genome editing tools can be used for functional testing in non-model species., Competing Interests: 8Conflict of Interest Statement The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
- Published
- 2020
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20. Co-evolving wing spots and mating displays are genetically separable traits in Drosophila.
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Massey JH, Rice GR, Firdaus AS, Chen CY, Yeh SD, Stern DL, and Wittkopp PJ
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- Animals, Female, Genes, X-Linked, Male, Sex Characteristics, Wings, Animal, Biological Coevolution, Drosophila genetics, Evolution, Molecular, Pigmentation genetics, Sexual Behavior, Animal
- Abstract
The evolution of sexual traits often involves correlated changes in morphology and behavior. For example, in Drosophila, divergent mating displays are often accompanied by divergent pigment patterns. To better understand how such traits co-evolve, we investigated the genetic basis of correlated divergence in wing pigmentation and mating display between the sibling species Drosophila elegans and Drosophila gunungcola. Drosophila elegans males have an area of black pigment on their wings known as a wing spot and appear to display this spot to females by extending their wings laterally during courtship. By contrast, D. gunungcola lost both of these traits. Using Multiplexed Shotgun Genotyping (MSG), we identified a ∼440 kb region on the X chromosome that behaves like a genetic switch controlling the presence or absence of male-specific wing spots. This region includes the candidate gene optomotor-blind (omb), which plays a critical role in patterning the Drosophila wing. The genetic basis of divergent wing display is more complex, with at least two loci on the X chromosome and two loci on autosomes contributing to its evolution. Introgressing the X-linked region affecting wing spot development from D. gunungcola into D. elegans reduced pigmentation in the wing spots but did not affect the wing display, indicating that these are genetically separable traits. Consistent with this observation, broader sampling of wild D. gunungcola populations confirmed that the wing spot and wing display are evolving independently: some D. gunungcola males performed wing displays similar to D. elegans despite lacking wing spots. These data suggest that correlated selection pressures rather than physical linkage or pleiotropy are responsible for the coevolution of these morphological and behavioral traits. They also suggest that the change in morphology evolved prior to the change in behavior., (© 2020 The Authors. Evolution © 2020 The Society for the Study of Evolution.)
- Published
- 2020
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21. Empirical measures of mutational effects define neutral models of regulatory evolution in Saccharomyces cerevisiae .
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Hodgins-Davis A, Duveau F, Walker EA, and Wittkopp PJ
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- Evolution, Molecular, Gene Expression genetics, Genome, Fungal genetics, Phenotype, Selection, Genetic genetics, Point Mutation genetics, Saccharomyces cerevisiae genetics
- Abstract
Understanding how phenotypes evolve requires disentangling the effects of mutation generating new variation from the effects of selection filtering it. Tests for selection frequently assume that mutation introduces phenotypic variation symmetrically around the population mean, yet few studies have tested this assumption by deeply sampling the distributions of mutational effects for particular traits. Here, we examine distributions of mutational effects for gene expression in the budding yeast Saccharomyces cerevisiae by measuring the effects of thousands of point mutations introduced randomly throughout the genome. We find that the distributions of mutational effects differ for the 10 genes surveyed and are inconsistent with normality. For example, all 10 distributions of mutational effects included more mutations with large effects than expected for normally distributed phenotypes. In addition, some genes also showed asymmetries in their distribution of mutational effects, with new mutations more likely to increase than decrease the gene's expression or vice versa. Neutral models of regulatory evolution that take these empirically determined distributions into account suggest that neutral processes may explain more expression variation within natural populations than currently appreciated., Competing Interests: The authors declare no competing interest.
- Published
- 2019
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22. The yellow gene influences Drosophila male mating success through sex comb melanization.
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Massey JH, Chung D, Siwanowicz I, Stern DL, and Wittkopp PJ
- Subjects
- Animals, Biological Evolution, Biomechanical Phenomena, Copulation physiology, Courtship, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Drosophila Proteins deficiency, Drosophila Proteins metabolism, Drosophila melanogaster anatomy & histology, Drosophila melanogaster metabolism, Extremities anatomy & histology, Female, Gene Expression Regulation, Male, Nerve Tissue Proteins genetics, Nerve Tissue Proteins metabolism, Transcription Factors genetics, Transcription Factors metabolism, Drosophila Proteins genetics, Drosophila melanogaster genetics, Mating Preference, Animal physiology, Pigmentation genetics
- Abstract
Drosophila melanogaster males perform a series of courtship behaviors that, when successful, result in copulation with a female. For over a century, mutations in the yellow gene, named for its effects on pigmentation, have been known to reduce male mating success. Prior work has suggested that yellow influences mating behavior through effects on wing extension, song, and/or courtship vigor. Here, we rule out these explanations, as well as effects on the nervous system more generally, and find instead that the effects of yellow on male mating success are mediated by its effects on pigmentation of male-specific leg structures called sex combs. Loss of yellow expression in these modified bristles reduces their melanization, which changes their structure and causes difficulty grasping females prior to copulation. These data illustrate why the mechanical properties of anatomy, not just neural circuitry, must be considered to fully understand the development and evolution of behavior., Competing Interests: JM, DC, IS, DS No competing interests declared, PW Senior editor, eLife, (© 2019, Massey et al.)
- Published
- 2019
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23. Compensatory trans -regulatory alleles minimizing variation in TDH3 expression are common within Saccharomyces cerevisiae .
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Metzger BPH and Wittkopp PJ
- Abstract
Heritable variation in gene expression is common within species. Much of this variation is due to genetic differences outside of the gene with altered expression and is trans -acting. This trans -regulatory variation is often polygenic, with individual variants typically having small effects, making the genetic architecture and evolution of trans -regulatory variation challenging to study. Consequently, key questions about trans -regulatory variation remain, including the variability of trans -regulatory variation within a species, how selection affects trans -regulatory variation, and how trans -regulatory variants are distributed throughout the genome and within a species. To address these questions, we isolated and measured trans -regulatory differences affecting TDH3 promoter activity among 56 strains of Saccharomyces cerevisiae , finding that trans -regulatory backgrounds varied approximately twofold in their effects on TDH3 promoter activity. Comparing this variation to neutral models of trans -regulatory evolution based on empirical measures of mutational effects revealed that despite this variability in the effects of trans -regulatory backgrounds, stabilizing selection has constrained trans- regulatory differences within this species. Using a powerful quantitative trait locus mapping method, we identified ∼100 trans -acting expression quantitative trait locus in each of three crosses to a common reference strain, indicating that regulatory variation is more polygenic than previous studies have suggested. Loci altering expression were located throughout the genome, and many loci were strain specific. This distribution and prevalence of alleles is consistent with recent theories about the genetic architecture of complex traits. In all mapping experiments, the nonreference strain alleles increased and decreased TDH3 promoter activity with similar frequencies, suggesting that stabilizing selection maintained many trans -acting variants with opposing effects. This variation may provide the raw material for compensatory evolution and larger scale regulatory rewiring observed in developmental systems drift among species., Competing Interests: 6The authors declare no conflicts of interest., (© 2019 The Author(s). Evolution Letters published by Wiley Periodicals, Inc. on behalf of Society for the Study of Evolution (SSE) and European Society for Evolutionary Biology (ESEB).)
- Published
- 2019
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24. Pleiotropic Effects of ebony and tan on Pigmentation and Cuticular Hydrocarbon Composition in Drosophila melanogaster .
- Author
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Massey JH, Akiyama N, Bien T, Dreisewerd K, Wittkopp PJ, Yew JY, and Takahashi A
- Abstract
Pleiotropic genes are genes that affect more than one trait. For example, many genes required for pigmentation in the fruit fly Drosophila melanogaster also affect traits such as circadian rhythms, vision, and mating behavior. Here, we present evidence that two pigmentation genes, ebony and tan , which encode enzymes catalyzing reciprocal reactions in the melanin biosynthesis pathway, also affect cuticular hydrocarbon (CHC) composition in D. melanogaster females. More specifically, we report that ebony loss-of-function mutants have a CHC profile that is biased toward long (>25C) chain CHCs, whereas tan loss-of-function mutants have a CHC profile that is biased toward short (<25C) chain CHCs. Moreover, pharmacological inhibition of dopamine synthesis, a key step in the melanin synthesis pathway, reversed the changes in CHC composition seen in ebony mutants, making the CHC profiles similar to those seen in tan mutants. These observations suggest that genetic variation affecting ebony and/or tan activity might cause correlated changes in pigmentation and CHC composition in natural populations. We tested this possibility using the Drosophila Genetic Reference Panel (DGRP) and found that CHC composition covaried with pigmentation as well as levels of ebony and tan expression in newly eclosed adults in a manner consistent with the ebony and tan mutant phenotypes. These data suggest that the pleiotropic effects of ebony and tan might contribute to covariation of pigmentation and CHC profiles in Drosophila .
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- 2019
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25. Corrigendum: Tempo and mode of regulatory evolution in Drosophila .
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Coolon JD, McManus CJ, Stevenson KR, Graveley BR, and Wittkopp PJ
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- 2018
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26. Fitness effects of altering gene expression noise in Saccharomyces cerevisiae .
- Author
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Duveau F, Hodgins-Davis A, Metzger BP, Yang B, Tryban S, Walker EA, Lybrook T, and Wittkopp PJ
- Subjects
- Gene Expression Regulation, Fungal genetics, Genotype, Single-Cell Analysis, Genetic Fitness genetics, Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) genetics, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins genetics, Selection, Genetic
- Abstract
Gene expression noise is an evolvable property of biological systems that describes differences in expression among genetically identical cells in the same environment. Prior work has shown that expression noise is heritable and can be shaped by selection, but the impact of variation in expression noise on organismal fitness has proven difficult to measure. Here, we quantify the fitness effects of altering expression noise for the TDH3 gene in Saccharomyces cerevisiae . We show that increases in expression noise can be deleterious or beneficial depending on the difference between the average expression level of a genotype and the expression level maximizing fitness. We also show that a simple model relating single-cell expression levels to population growth produces patterns consistent with our empirical data. We use this model to explore a broad range of average expression levels and expression noise, providing additional insight into the fitness effects of variation in expression noise., Competing Interests: FD, AH, BM, BY, ST, EW, TL No competing interests declared, PW Senior editor, eLife, (© 2018, Duveau et al.)
- Published
- 2018
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27. Effects of mutation and selection on plasticity of a promoter activity in Saccharomyces cerevisiae .
- Author
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Duveau F, Yuan DC, Metzger BPH, Hodgins-Davis A, and Wittkopp PJ
- Subjects
- Galactose metabolism, Glucose metabolism, Alleles, Gene Expression Regulation, Fungal, Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) biosynthesis, Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) genetics, Point Mutation, Response Elements, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins biosynthesis, Saccharomyces cerevisiae Proteins genetics
- Abstract
Phenotypic plasticity is an evolvable property of biological systems that can arise from environment-specific regulation of gene expression. To better understand the evolutionary and molecular mechanisms that give rise to plasticity in gene expression, we quantified the effects of 235 single-nucleotide mutations in the Saccharomyces cerevisiae TDH3 promoter ( P
TDH3 ) on the activity of this promoter in media containing glucose, galactose, or glycerol as a carbon source. We found that the distributions of mutational effects differed among environments because many mutations altered the plastic response exhibited by the wild-type allele. Comparing the effects of these mutations with the effects of 30 PTDH3 polymorphisms on expression plasticity in the same environments provided evidence of natural selection acting to prevent the plastic response in PTDH3 activity between glucose and galactose from becoming larger. The largest changes in expression plasticity were observed between fermentable (glucose or galactose) and nonfermentable (glycerol) carbon sources and were caused by mutations located in the RAP1 and GCR1 transcription factor binding sites. Mutations altered expression plasticity most frequently between the two fermentable environments, with mutations causing significant changes in plasticity between glucose and galactose distributed throughout the promoter, suggesting they might affect chromatin structure. Taken together, these results provide insight into the molecular mechanisms underlying gene-by-environment interactions affecting gene expression as well as the evolutionary dynamics affecting natural variation in plasticity of gene expression., Competing Interests: The authors declare no conflict of interest.- Published
- 2017
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28. Fitness Effects of Cis-Regulatory Variants in the Saccharomyces cerevisiae TDH3 Promoter.
- Author
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Duveau F, Toubiana W, and Wittkopp PJ
- Subjects
- Alleles, Gene Expression genetics, Gene Expression Regulation, Fungal genetics, Genetic Fitness genetics, Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) metabolism, Mutation, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins metabolism, Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) genetics, Promoter Regions, Genetic genetics, Saccharomyces cerevisiae Proteins genetics
- Abstract
Variation in gene expression is widespread within and between species, but fitness consequences of this variation are generally unknown. Here, we use mutations in the Saccharomyces cerevisiae TDH3 promoter to assess how changes in TDH3 expression affect cell growth. From these data, we predict the fitness consequences of de novo mutations and natural polymorphisms in the TDH3 promoter. Nearly all mutations and polymorphisms in the TDH3 promoter were found to have no significant effect on fitness in the environment assayed, suggesting that the wild-type allele of this promoter is robust to the effects of most new cis-regulatory mutations., (© The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2017
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29. Structure of the Transcriptional Regulatory Network Correlates with Regulatory Divergence in Drosophila.
- Author
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Yang B and Wittkopp PJ
- Subjects
- Animals, Biological Evolution, Drosophila genetics, Drosophila Proteins genetics, Evolution, Molecular, Gene Expression Regulation genetics, Genes, Insect, Genetic Association Studies, Genotype, Phenotype, RNA, Messenger genetics, Species Specificity, Gene Regulatory Networks genetics, Transcription Factors genetics
- Abstract
Transcriptional control of gene expression is regulated by biochemical interactions between cis-regulatory DNA sequences and trans-acting factors that form complex regulatory networks. Genetic changes affecting both cis- and trans-acting sequences in these networks have been shown to alter patterns of gene expression as well as higher-order organismal phenotypes. Here, we investigate how the structure of these regulatory networks relates to patterns of polymorphism and divergence in gene expression. To do this, we compared a transcriptional regulatory network inferred for Drosophila melanogaster to differences in gene regulation observed between two strains of D. melanogaster as well as between two pairs of closely related species: Drosophila sechellia and Drosophila simulans, and D. simulans and D. melanogaster. We found that the number of transcription factors predicted to directly regulate a gene ("in-degree") was negatively correlated with divergence in both gene expression (mRNA abundance) and cis-regulation. This observation suggests that the number of transcription factors directly regulating a gene's expression affects the conservation of cis-regulation and gene expression over evolutionary time. We also tested the hypothesis that transcription factors regulating more target genes (higher "out-degree") are less likely to evolve changes in their cis-regulation and expression (presumably due to increased pleiotropy), but found little support for this predicted relationship. Taken together, these data show how the architecture of regulatory networks can influence regulatory evolution., (© The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2017
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30. Evolutionary Dynamics of Regulatory Changes Underlying Gene Expression Divergence among Saccharomyces Species.
- Author
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Metzger BPH, Wittkopp PJ, and Coolon JD
- Subjects
- Alleles, Gene Expression Regulation, Fungal, Saccharomyces genetics, Species Specificity, Evolution, Molecular, Genetic Variation, Regulatory Sequences, Nucleic Acid genetics
- Abstract
Heritable changes in gene expression are important contributors to phenotypic differences within and between species and are caused by mutations in cis-regulatory elements and trans-regulatory factors. Although previous work has suggested that cis-regulatory differences preferentially accumulate with time, technical restrictions to closely related species and limited comparisons have made this observation difficult to test. To address this problem, we used allele-specific RNA-seq data from Saccharomyces species and hybrids to expand both the evolutionary timescale and number of species in which the evolution of regulatory divergence has been investigated. We find that as sequence divergence increases, cis-regulatory differences do indeed become the dominant type of regulatory difference between species, ultimately becoming a better predictor of expression divergence than trans-regulatory divergence. When both cis- and trans-regulatory differences accumulate for the same gene, they more often have effects in opposite directions than in the same direction, indicating widespread compensatory changes underlying the evolution of gene expression. The frequency of compensatory changes within and between species and the magnitude of effect for the underlying cis- and trans-regulatory differences suggests that compensatory changes accumulate primarily due to selection against divergence in gene expression as a result of weak stabilizing selection on gene expression levels. These results show that cis-regulatory differences and compensatory changes in regulation play increasingly important roles in the evolution of gene expression as time increases., (© The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)
- Published
- 2017
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31. Genetic basis of octanoic acid resistance in Drosophila sechellia: functional analysis of a fine-mapped region.
- Author
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Andrade López JM, Lanno SM, Auerbach JM, Moskowitz EC, Sligar LA, Wittkopp PJ, and Coolon JD
- Subjects
- Animals, Gene Knockdown Techniques, RNA Interference, Salivary Glands, Seychelles, Species Specificity, Caprylates, Drosophila genetics, Drosophila Proteins genetics
- Abstract
Drosophila sechellia is a species of fruit fly endemic to the Seychelles islands. Unlike its generalist sister species, D. sechellia has evolved to be a specialist on the host plant Morinda citrifolia. This specialization is interesting because the plant's fruit contains secondary defence compounds, primarily octanoic acid (OA), that are lethal to most other Drosophilids. Although ecological and behavioural adaptations to this toxic fruit are known, the genetic basis for evolutionary changes in OA resistance is not. Prior work showed that a genomic region on chromosome 3R containing 18 genes has the greatest contribution to differences in OA resistance between D. sechellia and D. simulans. To determine which gene(s) in this region might be involved in the evolutionary change in OA resistance, we knocked down expression of each gene in this region in D. melanogaster with RNA interference (RNAi) (i) ubiquitously throughout development, (ii) during only the adult stage and (iii) within specific tissues. We identified three neighbouring genes in the Osiris family, Osiris 6 (Osi6), Osi7 and Osi8, that led to decreased OA resistance when ubiquitously knocked down. Tissue-specific RNAi, however, showed that decreasing expression of Osi6 and Osi7 specifically in the fat body and/or salivary glands increased OA resistance. Gene expression analyses of Osi6 and Osi7 revealed that while standing levels of expression are higher in D. sechellia, Osi6 expression is significantly downregulated in salivary glands in response to OA exposure, suggesting that evolved tissue-specific environmental plasticity of Osi6 expression may be responsible for OA resistance in D. sechellia., (© 2016 John Wiley & Sons Ltd.)
- Published
- 2017
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32. Tools and strategies for scarless allele replacement in Drosophila using CRISPR/Cas9.
- Author
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Lamb AM, Walker EA, and Wittkopp PJ
- Subjects
- Alleles, Animals, Animals, Genetically Modified, Drosophila Proteins antagonists & inhibitors, Drosophila Proteins genetics, Drosophila melanogaster growth & development, Mutation, CRISPR-Cas Systems, Drosophila Proteins metabolism, Drosophila melanogaster genetics, Gene Editing methods
- Abstract
Genome editing via the CRISPR/Cas9 RNA-guided nuclease system has opened up exciting possibilities for genetic analysis. However, technical challenges associated with homology-directed repair have proven to be roadblocks for producing changes in the absence of unwanted, secondary mutations commonly known as "scars." To address these issues, we developed a 2-stage, marker-assisted strategy to facilitate precise, "scarless" edits in Drosophila with a minimal requirement for molecular screening. Using this method, we modified 2 base pairs in a gene of interest without altering the final sequence of the CRISPR cut sites. We executed this 2-stage allele swap using a novel transformation marker that drives expression in the pupal wings, which can be screened for in the presence of common eye-expressing reporters. The tools we developed can be used to make a single change or a series of allelic substitutions in a region of interest in any D. melanogaster genetic background as well as in other Drosophila species.
- Published
- 2017
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33. Potential Direct Regulators of the Drosophila yellow Gene Identified by Yeast One-Hybrid and RNAi Screens.
- Author
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Kalay G, Lusk R, Dome M, Hens K, Deplancke B, and Wittkopp PJ
- Subjects
- Animals, Drosophila metabolism, Ecdysone metabolism, Enhancer Elements, Genetic, Genetic Testing, Mutation, Phenotype, Protein Binding, Transcription Factors genetics, Transcription Factors metabolism, Drosophila genetics, Drosophila Proteins genetics, Gene Expression Regulation, Genetic Association Studies methods, Pigmentation genetics, RNA Interference, Two-Hybrid System Techniques
- Abstract
The regulation of gene expression controls development, and changes in this regulation often contribute to phenotypic evolution. Drosophila pigmentation is a model system for studying evolutionary changes in gene regulation, with differences in expression of pigmentation genes such as yellow that correlate with divergent pigment patterns among species shown to be caused by changes in cis- and trans-regulation. Currently, much more is known about the cis-regulatory component of divergent yellow expression than the trans-regulatory component, in part because very few trans-acting regulators of yellow expression have been identified. This study aims to improve our understanding of the trans-acting control of yellow expression by combining yeast-one-hybrid and RNAi screens for transcription factors binding to yellow cis-regulatory sequences and affecting abdominal pigmentation in adults, respectively. Of the 670 transcription factors included in the yeast-one-hybrid screen, 45 showed evidence of binding to one or more sequence fragments tested from the 5' intergenic and intronic yellow sequences from D. melanogaster, D. pseudoobscura, and D. willistoni, suggesting that they might be direct regulators of yellow expression. Of the 670 transcription factors included in the yeast-one-hybrid screen, plus another TF previously shown to be genetically upstream of yellow, 125 were also tested using RNAi, and 32 showed altered abdominal pigmentation. Nine transcription factors were identified in both screens, including four nuclear receptors related to ecdysone signaling (Hr78, Hr38, Hr46, and Eip78C). This finding suggests that yellow expression might be directly controlled by nuclear receptors influenced by ecdysone during early pupal development when adult pigmentation is forming., (Copyright © 2016 Kalay et al.)
- Published
- 2016
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34. Sensitivity of Allelic Divergence to Genomic Position: Lessons from the Drosophila tan Gene.
- Author
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John AV, Sramkoski LL, Walker EA, Cooley AM, and Wittkopp PJ
- Subjects
- Animals, Animals, Genetically Modified, Chromosomal Proteins, Non-Histone biosynthesis, DNA-Binding Proteins biosynthesis, Drosophila Proteins biosynthesis, Genome, Insect, Genomics, Integrases genetics, Phenotype, Species Specificity, Alleles, Chromosomal Proteins, Non-Histone genetics, DNA-Binding Proteins genetics, Drosophila genetics, Drosophila Proteins genetics, Transgenes genetics
- Abstract
To identify genetic variants underlying changes in phenotypes within and between species, researchers often utilize transgenic animals to compare the function of alleles in different genetic backgrounds. In Drosophila, targeted integration mediated by the ΦC31 integrase allows activity of alternative alleles to be compared at the same genomic location. By using the same insertion site for each transgene, position effects are generally assumed to be controlled for because both alleles are surrounded by the same genomic context. Here, we test this assumption by comparing the activity of tan alleles from two Drosophila species, D. americana and D. novamexicana, at five different genomic locations in D. melanogaster We found that the relative effects of these alleles varied among insertion sites, with no difference in activity observed between them at two sites. One of these sites simply silenced both transgenes, but the other allowed expression of both alleles that was sufficient to rescue a mutant phenotype yet failed to reveal the functional differences between the two alleles. These results suggest that more than one insertion site should be used when comparing the activity of transgenes because failing to do so could cause functional differences between alleles to go undetected., (Copyright © 2016 John et al.)
- Published
- 2016
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35. Contrasting Frequencies and Effects of cis- and trans-Regulatory Mutations Affecting Gene Expression.
- Author
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Metzger BP, Duveau F, Yuan DC, Tryban S, Yang B, and Wittkopp PJ
- Subjects
- Alleles, Base Sequence, Evolution, Molecular, Gene Expression, Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) metabolism, Mutation, Promoter Regions, Genetic, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins metabolism, Selection, Genetic, Gene Expression Regulation, Fungal, Genetic Complementation Test methods, Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) genetics, Regulatory Sequences, Nucleic Acid, Saccharomyces cerevisiae Proteins genetics
- Abstract
Heritable differences in gene expression are caused by mutations in DNA sequences encoding cis-regulatory elements and trans-regulatory factors. These two classes of regulatory change differ in their relative contributions to expression differences in natural populations because of the combined effects of mutation and natural selection. Here, we investigate how new mutations create the regulatory variation upon which natural selection acts by quantifying the frequencies and effects of hundreds of new cis- and trans-acting mutations altering activity of the TDH3 promoter in the yeast Saccharomyces cerevisiae in the absence of natural selection. We find that cis-regulatory mutations have larger effects on expression than trans-regulatory mutations and that while trans-regulatory mutations are more common overall, cis- and trans-regulatory changes in expression are equally abundant when only the largest changes in expression are considered. In addition, we find that cis-regulatory mutations are skewed toward decreased expression while trans-regulatory mutations are skewed toward increased expression. We also measure the effects of cis- and trans-regulatory mutations on the variability in gene expression among genetically identical cells, a property of gene expression known as expression noise, finding that trans-regulatory mutations are much more likely to decrease expression noise than cis-regulatory mutations. Because new mutations are the raw material upon which natural selection acts, these differences in the frequencies and effects of cis- and trans-regulatory mutations should be considered in models of regulatory evolution., (© The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2016
- Full Text
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36. The Genetic Basis of Pigmentation Differences Within and Between Drosophila Species.
- Author
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Massey JH and Wittkopp PJ
- Subjects
- Abdomen physiology, Animals, Organ Specificity genetics, Pupa genetics, Thorax physiology, Wings, Animal physiology, Drosophila genetics, Pigmentation genetics
- Abstract
In Drosophila, as well as in many other plants and animals, pigmentation is highly variable both within and between species. This variability, combined with powerful genetic and transgenic tools as well as knowledge of how pigment patterns are formed biochemically and developmentally, has made Drosophila pigmentation a premier system for investigating the genetic and molecular mechanisms responsible for phenotypic evolution. In this chapter, we review and synthesize findings from a rapidly growing body of case studies examining the genetic basis of pigmentation differences in the abdomen, thorax, wings, and pupal cases within and between Drosophila species. A core set of genes, including genes required for pigment synthesis (eg, yellow, ebony, tan, Dat) as well as developmental regulators of these genes (eg, bab1, bab2, omb, Dll, and wg), emerge as the primary sources of this variation, with most genes having been shown to contribute to pigmentation differences both within and between species. In cases where specific genetic changes contributing to pigmentation divergence were identified in these genes, the changes were always located in noncoding sequences and affected cis-regulatory activity. We conclude this chapter by discussing these and other lessons learned from evolutionary genetic studies of Drosophila pigmentation and identify topics we think should be the focus of future work with this model system., (© 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
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37. Molecular Mechanisms and Evolutionary Processes Contributing to Accelerated Divergence of Gene Expression on the Drosophila X Chromosome.
- Author
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Coolon JD, Stevenson KR, McManus CJ, Yang B, Graveley BR, and Wittkopp PJ
- Subjects
- Animals, Base Sequence, Female, Genes, X-Linked, Genetic Variation, Male, Regulatory Sequences, Nucleic Acid genetics, Transcription Factors metabolism, Biological Evolution, Drosophila melanogaster genetics, Gene Expression Regulation, X Chromosome genetics
- Abstract
In species with a heterogametic sex, population genetics theory predicts that DNA sequences on the X chromosome can evolve faster than comparable sequences on autosomes. Both neutral and nonneutral evolutionary processes can generate this pattern. Complex traits like gene expression are not predicted to have accelerated evolution by these theories, yet a "faster-X" pattern of gene expression divergence has recently been reported for both Drosophila and mammals. Here, we test the hypothesis that accelerated adaptive evolution of cis-regulatory sequences on the X chromosome is responsible for this pattern by comparing the relative contributions of cis- and trans-regulatory changes to patterns of faster-X expression divergence observed between strains and species of Drosophila with a range of divergence times. We find support for this hypothesis, especially among male-biased genes, when comparing different species. However, we also find evidence that trans-regulatory differences contribute to a faster-X pattern of expression divergence both within and between species. This contribution is surprising because trans-acting regulators of X-linked genes are generally assumed to be randomly distributed throughout the genome. We found, however, that X-linked transcription factors appear to preferentially regulate expression of X-linked genes, providing a potential mechanistic explanation for this result. The contribution of trans-regulatory variation to faster-X expression divergence was larger within than between species, suggesting that it is more likely to result from neutral processes than positive selection. These data show how accelerated evolution of both coding and noncoding sequences on the X chromosome can lead to accelerated expression divergence on the X chromosome relative to autosomes., (© The Author 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2015
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38. Selection on noise constrains variation in a eukaryotic promoter.
- Author
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Metzger BP, Yuan DC, Gruber JD, Duveau F, and Wittkopp PJ
- Subjects
- Evolution, Molecular, Gene Expression Regulation, Fungal genetics, Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) genetics, Mutation genetics, Phenotype, Saccharomyces cerevisiae Proteins genetics, Polymorphism, Genetic genetics, Promoter Regions, Genetic genetics, Saccharomyces cerevisiae genetics, Selection, Genetic genetics
- Abstract
Genetic variation segregating within a species reflects the combined activities of mutation, selection, and genetic drift. In the absence of selection, polymorphisms are expected to be a random subset of new mutations; thus, comparing the effects of polymorphisms and new mutations provides a test for selection. When evidence of selection exists, such comparisons can identify properties of mutations that are most likely to persist in natural populations. Here we investigate how mutation and selection have shaped variation in a cis-regulatory sequence controlling gene expression by empirically determining the effects of polymorphisms segregating in the TDH3 promoter among 85 strains of Saccharomyces cerevisiae and comparing their effects to a distribution of mutational effects defined by 236 point mutations in the same promoter. Surprisingly, we find that selection on expression noise (that is, variability in expression among genetically identical cells) appears to have had a greater impact on sequence variation in the TDH3 promoter than selection on mean expression level. This is not necessarily because variation in expression noise impacts fitness more than variation in mean expression level, but rather because of differences in the distributions of mutational effects for these two phenotypes. This study shows how systematically examining the effects of new mutations can enrich our understanding of evolutionary mechanisms. It also provides rare empirical evidence of selection acting on expression noise.
- Published
- 2015
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39. The significance and scope of evolutionary developmental biology: a vision for the 21st century.
- Author
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Moczek AP, Sears KE, Stollewerk A, Wittkopp PJ, Diggle P, Dworkin I, Ledon-Rettig C, Matus DQ, Roth S, Abouheif E, Brown FD, Chiu CH, Cohen CS, Tomaso AW, Gilbert SF, Hall B, Love AC, Lyons DC, Sanger TJ, Smith J, Specht C, Vallejo-Marin M, and Extavour CG
- Subjects
- Animals, Gene Regulatory Networks, Humans, Biological Evolution, Developmental Biology education, Developmental Biology trends, Genetics education, Genetics trends
- Abstract
Evolutionary developmental biology (evo-devo) has undergone dramatic transformations since its emergence as a distinct discipline. This paper aims to highlight the scope, power, and future promise of evo-devo to transform and unify diverse aspects of biology. We articulate key questions at the core of eleven biological disciplines-from Evolution, Development, Paleontology, and Neurobiology to Cellular and Molecular Biology, Quantitative Genetics, Human Diseases, Ecology, Agriculture and Science Education, and lastly, Evolutionary Developmental Biology itself-and discuss why evo-devo is uniquely situated to substantially improve our ability to find meaningful answers to these fundamental questions. We posit that the tools, concepts, and ways of thinking developed by evo-devo have profound potential to advance, integrate, and unify biological sciences as well as inform policy decisions and illuminate science education. We look to the next generation of evolutionary developmental biologists to help shape this process as we confront the scientific challenges of the 21st century., (© 2015 Wiley Periodicals, Inc.)
- Published
- 2015
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40. Evolution of splicing regulatory networks in Drosophila.
- Author
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McManus CJ, Coolon JD, Eipper-Mains J, Wittkopp PJ, and Graveley BR
- Subjects
- Animals, Base Sequence, Models, Genetic, Molecular Sequence Data, Species Specificity, Alternative Splicing, Drosophila genetics, Evolution, Molecular, Gene Regulatory Networks
- Abstract
The proteome expanding effects of alternative pre-mRNA splicing have had a profound impact on eukaryotic evolution. The events that create this diversity can be placed into four major classes: exon skipping, intron retention, alternative 5' splice sites, and alternative 3' splice sites. Although the regulatory mechanisms and evolutionary pressures among alternative splicing classes clearly differ, how these differences affect the evolution of splicing regulation remains poorly characterized. We used RNA-seq to investigate splicing differences in D. simulans, D. sechellia, and three strains of D. melanogaster. Regulation of exon skipping and tandem alternative 3' splice sites (NAGNAGs) were more divergent than other splicing classes. Splicing regulation was most divergent in frame-preserving events and events in noncoding regions. We further determined the contributions of cis- and trans-acting changes in splicing regulatory networks by comparing allele-specific splicing in F1 interspecific hybrids, because differences in allele-specific splicing reflect changes in cis-regulatory element activity. We find that species-specific differences in intron retention and alternative splice site usage are primarily attributable to changes in cis-regulatory elements (median ∼80% cis), whereas species-specific exon skipping differences are driven by both cis- and trans-regulatory divergence (median ∼50% cis). These results help define the mechanisms and constraints that influence splicing regulatory evolution and show that networks regulating the four major classes of alternative splicing diverge through different genetic mechanisms. We propose a model in which differences in regulatory network architecture among classes of alternative splicing affect the evolution of splicing regulation.
- Published
- 2014
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41. Tempo and mode of regulatory evolution in Drosophila.
- Author
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Coolon JD, McManus CJ, Stevenson KR, Graveley BR, and Wittkopp PJ
- Subjects
- Animals, Drosophila Proteins genetics, Drosophila Proteins metabolism, Time Factors, Transcription, Genetic, Drosophila genetics, Evolution, Molecular, Regulatory Sequences, Nucleic Acid genetics
- Abstract
Genetic changes affecting gene expression contribute to phenotypic divergence; thus, understanding how regulatory networks controlling gene expression change over time is critical for understanding evolution. Prior studies of expression differences within and between species have identified properties of regulatory divergence, but technical and biological differences among these studies make it difficult to assess the generality of these properties or to understand how regulatory changes accumulate with divergence time. Here, we address these issues by comparing gene expression among strains and species of Drosophila with a range of divergence times and use F1 hybrids to examine inheritance patterns and disentangle cis- and trans-regulatory changes. We find that the fixation of compensatory changes has caused the regulation of gene expression to diverge more rapidly than gene expression itself. Specifically, we observed that the proportion of genes with evidence of cis-regulatory divergence has increased more rapidly with divergence time than the proportion of genes with evidence of expression differences. Surprisingly, the amount of expression divergence explained by cis-regulatory changes did not increase steadily with divergence time, as was previously proposed. Rather, one species (Drosophila sechellia) showed an excess of cis-regulatory divergence that we argue most likely resulted from positive selection in this lineage. Taken together, this work reveals not only the rate at which gene expression evolves, but also the molecular and evolutionary mechanisms responsible for this evolution.
- Published
- 2014
- Full Text
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42. Mapping small effect mutations in Saccharomyces cerevisiae: impacts of experimental design and mutational properties.
- Author
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Duveau F, Metzger BP, Gruber JD, Mack K, Sood N, Brooks TE, and Wittkopp PJ
- Subjects
- Bacterial Proteins genetics, Bacterial Proteins metabolism, Genes, Reporter, Genetic Variation, High-Throughput Nucleotide Sequencing, Luminescent Proteins genetics, Luminescent Proteins metabolism, Mutagenesis, Site-Directed, Sequence Analysis, DNA, Chromosome Mapping, Research Design, Saccharomyces cerevisiae genetics
- Abstract
Genetic variants identified by mapping are biased toward large phenotypic effects because of methodologic challenges for detecting genetic variants with small phenotypic effects. Recently, bulk segregant analysis combined with next-generation sequencing (BSA-seq) was shown to be a powerful and cost-effective way to map small effect variants in natural populations. Here, we examine the power of BSA-seq for efficiently mapping small effect mutations isolated from a mutagenesis screen. Specifically, we determined the impact of segregant population size, intensity of phenotypic selection to collect segregants, number of mitotic generations between meiosis and sequencing, and average sequencing depth on power for mapping mutations with a range of effects on the phenotypic mean and standard deviation as well as relative fitness. We then used BSA-seq to map the mutations responsible for three ethyl methanesulfonate-induced mutant phenotypes in Saccharomyces cerevisiae. These mutants display small quantitative variation in the mean expression of a fluorescent reporter gene (-3%, +7%, and +10%). Using a genetic background with increased meiosis rate, a reliable mating type marker, and fluorescence-activated cell sorting to efficiently score large segregating populations and isolate cells with extreme phenotypes, we successfully mapped and functionally confirmed a single point mutation responsible for the mutant phenotype in all three cases. Our simulations and experimental data show that the effects of a causative site not only on the mean phenotype, but also on its standard deviation and relative fitness should be considered when mapping genetic variants in microorganisms such as yeast that require population growth steps for BSA-seq., (Copyright © 2014 Duveau et al.)
- Published
- 2014
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43. Effect of genetic variation in a Drosophila model of diabetes-associated misfolded human proinsulin.
- Author
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He BZ, Ludwig MZ, Dickerson DA, Barse L, Arun B, Vilhjálmsson BJ, Jiang P, Park SY, Tamarina NA, Selleck SB, Wittkopp PJ, Bell GI, and Kreitman M
- Subjects
- Alleles, Animals, Animals, Genetically Modified, Diabetes Mellitus metabolism, Disease Models, Animal, Drosophila Proteins chemistry, Drosophila Proteins genetics, Drosophila Proteins metabolism, Drosophila melanogaster, Epistasis, Genetic, Eye metabolism, Eye pathology, Female, Gene Expression, Gene Knockdown Techniques, Genome-Wide Association Study, Heparitin Sulfate biosynthesis, Humans, Introns, Male, Mutation, Phenotype, Proinsulin chemistry, Protein Folding, RNA Interference, Sulfotransferases chemistry, Sulfotransferases genetics, Sulfotransferases metabolism, Diabetes Mellitus genetics, Genetic Variation, Proinsulin genetics
- Abstract
The identification and validation of gene-gene interactions is a major challenge in human studies. Here, we explore an approach for studying epistasis in humans using a Drosophila melanogaster model of neonatal diabetes mellitus. Expression of the mutant preproinsulin (hINS(C96Y)) in the eye imaginal disc mimics the human disease: it activates conserved stress-response pathways and leads to cell death (reduction in eye area). Dominant-acting variants in wild-derived inbred lines from the Drosophila Genetics Reference Panel produce a continuous, highly heritable distribution of eye-degeneration phenotypes in a hINS(C96Y) background. A genome-wide association study (GWAS) in 154 sequenced lines identified a sharp peak on chromosome 3L, which mapped to a 400-bp linkage block within an intron of the gene sulfateless (sfl). RNAi knockdown of sfl enhanced the eye-degeneration phenotype in a mutant-hINS-dependent manner. RNAi against two additional genes in the heparan sulfate (HS) biosynthetic pathway (ttv and botv), in which sfl acts, also modified the eye phenotype in a hINS(C96Y)-dependent manner, strongly suggesting a novel link between HS-modified proteins and cellular responses to misfolded proteins. Finally, we evaluated allele-specific expression difference between the two major sfl-intronic haplotypes in heterozygtes. The results showed significant heterogeneity in marker-associated gene expression, thereby leaving the causal mutation(s) and its mechanism unidentified. In conclusion, the ability to create a model of human genetic disease, map a QTL by GWAS to a specific gene, and validate its contribution to disease with available genetic resources and the potential to experimentally link the variant to a molecular mechanism demonstrate the many advantages Drosophila holds in determining the genetic underpinnings of human disease.
- Published
- 2014
- Full Text
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44. The roles of cis- and trans-regulation in the evolution of regulatory incompatibilities and sexually dimorphic gene expression.
- Author
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Meiklejohn CD, Coolon JD, Hartl DL, and Wittkopp PJ
- Subjects
- Alleles, Animals, Biological Evolution, Female, Gene Expression Profiling, Hybridization, Genetic, Male, Models, Genetic, Organ Specificity, Sex Characteristics, Species Specificity, Drosophila genetics, Evolution, Molecular, Gene Expression Regulation, Genome, Insect, Regulatory Elements, Transcriptional
- Abstract
Evolutionary changes in gene expression underlie many aspects of phenotypic diversity within and among species. Understanding the genetic basis for evolved changes in gene expression is therefore an important component of a comprehensive understanding of the genetic basis of phenotypic evolution. Using interspecific introgression hybrids, we examined the genetic basis for divergence in genome-wide patterns of gene expression between Drosophila simulans and Drosophila mauritiana. We find that cis-regulatory and trans-regulatory divergences differ significantly in patterns of genetic architecture and evolution. The effects of cis-regulatory divergence are approximately additive in heterozygotes, quantitatively different between males and females, and well predicted by expression differences between the two parental species. In contrast, the effects of trans-regulatory divergence are associated with largely dominant introgressed alleles, have similar effects in the two sexes, and generate expression levels in hybrids outside the range of expression in both parental species. Although the effects of introgressed trans-regulatory alleles are similar in males and females, expression levels of the genes they regulate are sexually dimorphic between the parental D. simulans and D. mauritiana strains, suggesting that pure-species genotypes carry unlinked modifier alleles that increase sexual dimorphism in expression. Our results suggest that independent effects of cis-regulatory substitutions in males and females may favor their role in the evolution of sexually dimorphic phenotypes, and that trans-regulatory divergence is an important source of regulatory incompatibilities.
- Published
- 2014
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- View/download PDF
45. Sex-specific effects of cis-regulatory variants in Drosophila melanogaster.
- Author
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Coolon JD, Webb W, and Wittkopp PJ
- Subjects
- Animals, Chromosomes, Insect genetics, Drosophila Proteins metabolism, Female, Male, Sex Chromosomes genetics, Transcription, Genetic, Drosophila Proteins genetics, Drosophila melanogaster genetics, Epistasis, Genetic, Genomic Imprinting, Sex Characteristics
- Abstract
Sexual dimorphism at the level of gene expression is common and well documented, but much less is known about how different cis-regulatory alleles interact with the different trans-regulatory environments present in males and females. Here we show that sex-specific effects of cis-regulatory variants are common in Drosophila.
- Published
- 2013
- Full Text
- View/download PDF
46. Sources of bias in measures of allele-specific expression derived from RNA-sequence data aligned to a single reference genome.
- Author
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Stevenson KR, Coolon JD, and Wittkopp PJ
- Subjects
- Animals, Drosophila genetics, Exons, Alleles, Genome, Sequence Analysis, RNA
- Abstract
Background: RNA-seq can be used to measure allele-specific expression (ASE) by assigning sequence reads to individual alleles; however, relative ASE is systematically biased when sequence reads are aligned to a single reference genome. Aligning sequence reads to both parental genomes can eliminate this bias, but this approach is not always practical, especially for non-model organisms. To improve accuracy of ASE measured using a single reference genome, we identified properties of differentiating sites responsible for biased measures of relative ASE., Results: We found that clusters of differentiating sites prevented sequence reads from an alternate allele from aligning to the reference genome, causing a bias in relative ASE favoring the reference allele. This bias increased with greater sequence divergence between alleles. Increasing the number of mismatches allowed when aligning sequence reads to the reference genome and restricting analysis to genomic regions with fewer differentiating sites than the number of mismatches allowed almost completely eliminated this systematic bias. Accuracy of allelic abundance was increased further by excluding differentiating sites within sequence reads that could not be aligned uniquely within the genome (imperfect mappability) and reads that overlapped one or more insertions or deletions (indels) between alleles., Conclusions: After aligning sequence reads to a single reference genome, excluding differentiating sites with at least as many neighboring differentiating sites as the number of mismatches allowed, imperfect mappability, and/or an indel(s) nearby resulted in measures of allelic abundance comparable to those derived from aligning sequence reads to both parental genomes.
- Published
- 2013
- Full Text
- View/download PDF
47. Population genetics and a study of speciation using next-generation sequencing: an educational primer for use with "Patterns of transcriptome divergence in the male accessory gland of two closely related species of field crickets".
- Author
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Wittkopp PJ
- Subjects
- Animals, Male, Genetic Speciation, Gryllidae genetics, Semen metabolism, Transcriptome
- Abstract
Unlabelled: Understanding evidence for the genetic basis of reproductive isolation is imperative for supporting students' understanding of mechanisms of speciation in courses such as Genetics and Evolutionary Biology. An article by Andrés et al. in the February 2013 issue of GENETICS illustrates how advances in DNA sequencing are accelerating studies of population genetics in species with limited genetic and genomic resources. Andrés et al. use the latest sequencing technologies to systematically identify and characterize sites in the DNA that vary within, and have diverged between, species to explore speciation in crickets. This primer, coupled with that article, will help instructors introduce and reinforce important concepts in genetics and evolution while simultaneously introducing modern methodology in the undergraduate classroom. Related article in, Genetics: Andrés, J. A., E. L. Larson, S. M. Bogdanowicz, and R. G. Harrison, 2013 Patterns of transcriptome divergence in the male accessory gland of two closely related species of field crickets. Genetics 193: 501-513.
- Published
- 2013
- Full Text
- View/download PDF
48. Genomic imprinting absent in Drosophila melanogaster adult females.
- Author
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Coolon JD, Stevenson KR, McManus CJ, Graveley BR, and Wittkopp PJ
- Subjects
- Age Factors, Animals, Crosses, Genetic, Female, Gene Deletion, Gene Expression Regulation, Gene Frequency, Genes, Insect genetics, Male, Models, Biological, Multigene Family genetics, Sex Factors, Drosophila melanogaster genetics, Genomic Imprinting physiology
- Abstract
Genomic imprinting occurs when expression of an allele differs based on the sex of the parent that transmitted the allele. In D. melanogaster, imprinting can occur, but its impact on allelic expression genome-wide is unclear. Here, we search for imprinted genes in D. melanogaster using RNA-seq to compare allele-specific expression between pools of 7- to 10-day-old adult female progeny from reciprocal crosses. We identified 119 genes with allelic expression consistent with imprinting, and these genes showed significant clustering within the genome. Surprisingly, additional analysis of several of these genes showed that either genomic heterogeneity or high levels of intrinsic noise caused imprinting-like allelic expression. Consequently, our data provide no convincing evidence of imprinting for D. melanogaster genes in their native genomic context. Elucidating sources of false-positive signals for imprinting in allele-specific RNA-seq data, as done here, is critical given the growing popularity of this method for identifying imprinted genes., (Copyright © 2012 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
49. The ontogeny of color: developmental origins of divergent pigmentation in Drosophila americana and D. novamexicana.
- Author
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Cooley AM, Shefner L, McLaughlin WN, Stewart EE, and Wittkopp PJ
- Subjects
- Alleles, Animals, DNA-Binding Proteins biosynthesis, Drosophila metabolism, Drosophila Proteins biosynthesis, Phenotype, Quantitative Trait Loci, RNA, Messenger biosynthesis, Species Specificity, DNA-Binding Proteins genetics, Drosophila genetics, Drosophila Proteins genetics, Evolution, Molecular, Gene Expression Regulation genetics, Pigmentation genetics, RNA, Messenger genetics
- Abstract
Pigmentation is a model trait for evolutionary and developmental analysis that is particularly amenable to molecular investigation in the genus Drosophila. To better understand how this phenotype evolves, we examined divergent pigmentation and gene expression over developmental time in the dark-bodied D. americana and its light-bodied sister species D. novamexicana. Prior genetic analysis implicated two enzyme-encoding genes, tan and ebony, in pigmentation divergence between these species, but questions remain about the underlying molecular mechanisms. Here, we describe stages of pupal development in both species and use this staging to determine when pigmentation develops and diverges between D. americana and D. novamexicana. For the developmental stages encompassing pigment divergence, we compare mRNA expression of tan and ebony over time and between species. Finally, we use allele-specific expression assays to determine whether interspecific differences in mRNA abundance have a cis-regulatory basis and find evidence of cis-regulatory divergence for both tan and ebony. cis-regulatory divergence affecting tan had a small effect on mRNA abundance and was limited to a few developmental stages, yet previous data suggests that this divergence is likely to be biologically meaningful. Our study suggests that small and developmentally transient expression changes may contribute to phenotypic diversification more often than commonly appreciated. Recognizing the potential phenotypic impact of such changes is important for a scientific community increasingly focused on dissecting quantitative variation, but detecting these types of changes will be a major challenge to elucidating the molecular basis of complex traits., (© 2012 Wiley Periodicals, Inc.)
- Published
- 2012
- Full Text
- View/download PDF
50. Contrasting properties of gene-specific regulatory, coding, and copy number mutations in Saccharomyces cerevisiae: frequency, effects, and dominance.
- Author
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Gruber JD, Vogel K, Kalay G, and Wittkopp PJ
- Subjects
- Evolution, Molecular, Genes, Dominant, Genes, Recessive, Genotype, Haploidy, Heterozygote, Mutation Rate, Promoter Regions, Genetic, DNA Copy Number Variations genetics, Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) genetics, Mutation genetics, Open Reading Frames genetics, Regulatory Sequences, Nucleic Acid genetics, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins genetics
- Abstract
Genetic variation within and between species can be shaped by population-level processes and mutation; however, the relative impact of "survival of the fittest" and "arrival of the fittest" on phenotypic evolution remains unclear. Assessing the influence of mutation on evolution requires understanding the relative rates of different types of mutations and their genetic properties, yet little is known about the functional consequences of new mutations. Here, we examine the spectrum of mutations affecting a focal gene in Saccharomyces cerevisiae by characterizing 231 novel haploid genotypes with altered activity of a fluorescent reporter gene. 7% of these genotypes had a nonsynonymous mutation in the coding sequence for the fluorescent protein and were classified as "coding" mutants; 2% had a change in the S. cerevisiae TDH3 promoter sequence controlling expression of the fluorescent protein and were classified as "cis-regulatory" mutants; 10% contained two copies of the reporter gene and were classified as "copy number" mutants; and the remaining 81% showed altered fluorescence without a change in the reporter gene itself and were classified as "trans-acting" mutants. As a group, coding mutants had the strongest effect on reporter gene activity and always decreased it. By contrast, 50%-95% of the mutants in each of the other three classes increased gene activity, with mutants affecting copy number and cis-regulatory sequences having larger median effects on gene activity than trans-acting mutants. When made heterozygous in diploid cells, coding, cis-regulatory, and copy number mutant genotypes all had significant effects on gene activity, whereas 88% of the trans-acting mutants appeared to be recessive. These differences in the frequency, effects, and dominance among functional classes of mutations might help explain why some types of mutations are found to be segregating within or fixed between species more often than others., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2012
- Full Text
- View/download PDF
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