Comparison of testis transcriptome changes resulting from mutations in dany and representative meiotic arrest genes
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ABSTRACT:
ORGANISM(S): Drosophila melanogaster
PROVIDER: GSE76298 | GEO | 2016/07/28
SECONDARY ACCESSION(S): PRJNA306854
REPOSITORIES: GEO
Similar Datasets
Project description:The Drosophila gene dany is expressed specifically in spermatocytes and encodes a nuclear protein with similarity to the products of the genes distal antenna (dan) and distal antenna-related (danr). Loss of dany function results in male sterility. Mutant spermatocytes fail to differentiate similar as previously observed in mutants lacking tTAF and tMAC function. Microarray analyses were performed to characterize the effects of loss of dany function on gene expression in testis and for a comparison of the effects with those caused by mutations in the tTAF gene spermatocyte arrest (sa) and the tMAC genes always early (aly) and matotopetli (topi) RNA was isolated after testis dissection from adult males and used for the production of probes for microarray hybridization. Five different genotypes were used for testis dissection: control, dany mutant, sa mutant, aly mutant, topi mutant. The number of biological replicates per genotypes was between two and four.
2016-07-28 | E-GEOD-76298 | biostudies-arrayexpress
Project description:The role of different proteins, Always Early (Aly), Spermatocyte Arrest (Sa), Ubi-p63E (Magn) on the gene expression in spermatocyte differentation was assessed by microarray ABSTRACT: The ubiquitin proteasome system (UPS) regulates many biological pathways by posttranslationally ubiquitinating proteins for degradation. Although maintaining a dynamic balance between free ubiquitin and ubiquitinated proteins is key to UPS function, the mechanisms that regulate ubiquitin homeostasis in different tissues through development are not clear. Here we show that loss of function of Drosophila magellan (magn), the polyubiquitin Ubi-p63E, results in specifically meiotic arrest sterility in males. Expression of ubiquitin from magn/Ubi-p63E contributes predominantly to maintaining the free ubiquitin pool in testes. Function of magn/Ubip63E is required cell autonomously for proper meiotic chromatin condensation, cell cycle progression and spermatid differentiation. magn/Ubi-p63E mutant germ cells develop normally to the spermatocyte stage but arrest at the G2/M transition of meiosis I with lack of protein expression of key meiotic cell cycle regulators Boule and Cyclin B. Loss of function of magn/Ubi-p63E did not strongly affect the spermatocyte transcription program regulated by the tTAF and tMAC genes. Knocking down proteasome function specifically in spermatocytes caused a different meiotic arrest phenotype, suggesting that the magn/Ubi-p63E phenotype may not result from general defects in protein degradation. Our results suggest a conserved role of polyubiquitin genes in male meiosis and a potential mechanism leading to meiosis I maturation arrest. RNA was obtained from whole testes of flies with following mutations: 1) aly[2]/aly[5p], 2) sa[1]/sa[2], 3) magn[12c]/magn[23b] or magn[12c]/magn[12c];pSC2, 4) red[1], e[1] (WT). Each genotype had three biological replicates except magn which had four total biological replicates, two from magn[12c]/[23b] and two from [12c]/[12c]; pSC2
2013-07-13 | E-GEOD-48837 | biostudies-arrayexpress
Project description:The effect of Med22, spermatocyte arrest (sa) and always early (aly) loss of function on the gene expression in spermatocytes was assessed by microarray. Title: Recruitment of Mediator complex by cell type and stage-specific factors required for tissue-specific TAF dependent gene activation in an adult stem cell lineage
2015-11-07 | GSE74784 | GEO
Project description:The role of different proteins, Always Early (Aly), Spermatocyte Arrest (Sa), Ubi-p63E (Magn) on the gene expression in spermatocyte differentation was assessed by microarray ABSTRACT: The ubiquitin proteasome system (UPS) regulates many biological pathways by posttranslationally ubiquitinating proteins for degradation. Although maintaining a dynamic balance between free ubiquitin and ubiquitinated proteins is key to UPS function, the mechanisms that regulate ubiquitin homeostasis in different tissues through development are not clear. Here we show that loss of function of Drosophila magellan (magn), the polyubiquitin Ubi-p63E, results in specifically meiotic arrest sterility in males. Expression of ubiquitin from magn/Ubi-p63E contributes predominantly to maintaining the free ubiquitin pool in testes. Function of magn/Ubip63E is required cell autonomously for proper meiotic chromatin condensation, cell cycle progression and spermatid differentiation. magn/Ubi-p63E mutant germ cells develop normally to the spermatocyte stage but arrest at the G2/M transition of meiosis I with lack of protein expression of key meiotic cell cycle regulators Boule and Cyclin B. Loss of function of magn/Ubi-p63E did not strongly affect the spermatocyte transcription program regulated by the tTAF and tMAC genes. Knocking down proteasome function specifically in spermatocytes caused a different meiotic arrest phenotype, suggesting that the magn/Ubi-p63E phenotype may not result from general defects in protein degradation. Our results suggest a conserved role of polyubiquitin genes in male meiosis and a potential mechanism leading to meiosis I maturation arrest.
2013-07-13 | GSE48837 | GEO
Project description:The effect of Med22, spermatocyte arrest (sa) and always early (aly) loss of function on the gene expression in spermatocytes was assessed by microarray. Title: Recruitment of Mediator complex by cell type and stage-specific factors required for tissue-specific TAF dependent gene activation in an adult stem cell lineage RNA was obtained from whole testes of flies with following mutations: 1)bam-gal4: UAS-Med22RNAi (VDRC#104581). Three biological replicates were included. Experiments were performed at 25C.
2015-11-07 | E-GEOD-74784 | biostudies-arrayexpress
Project description:Animal germline development and fertility relies on paralogs of general transcription factors that recruit RNA polymerase II to ensure cell type-specific gene expression. It remains unclear whether gene expression processes downstream of such paralog-based transcription is distinct from that of canonical RNA polymerase II genes. In Drosophila, the testis-specific TBP-associated factors (tTAFs) activate over a thousand spermatocyte-specific gene promoters to enable meiosis and germ cell differentiation. Here we show that efficient termination of tTAF-activated transcription requires a testis-specific Polymerase Associated Factor 1 Complex (tPAF) composed of paralogs of canonical Polymerase Associated Factor 1 Complex (PAF1C) proteins. Defective transcription termination in tPAF mutants causes aberrant expression of hundreds of downstream genes due to read-in transcription, compromising cell type-specific gene expression in spermatocytes. Consistently, tPAF is required for the segregation of meiotic chromosomes, sperm individualisation, and male fertility. Comparative in vivo proximity labeling assays of tPAF and PAF1C showed tPAF-specific association with tTAF as well as connections to central RNA polymerase II termination factors. Our study uncovers transcription termination as a developmentally regulated process required for cell type-specific gene expression.
2024-04-13 | PXD049360 | Pride
Project description:Animal germline development and fertility relies on paralogs of general transcription factors that recruit RNA polymerase II to ensure cell type-specific gene expression. It remains unclear whether gene expression processes downstream of such paralog-based transcription is distinct from that of canonical RNA polymerase II genes. In Drosophila, the testis-specific TBP-associated factors (tTAFs) activate over a thousand spermatocyte-specific gene promoters to enable meiosis and germ cell differentiation. Here we show that efficient termination of tTAF-activated transcription requires a testis-specific Polymerase Associated Factor 1 Complex (tPAF) composed of paralogs of canonical Polymerase Associated Factor 1 Complex (PAF1C) proteins. Defective transcription termination in tPAF mutants causes aberrant expression of hundreds of downstream genes due to read-in transcription, compromising cell type-specific gene expression in spermatocytes. Consistently, tPAF is required for the segregation of meiotic chromosomes, sperm individualisation, and male fertility. Comparative in vivo proximity labeling assays of tPAF and PAF1C showed tPAF-specific association with tTAF as well as connections to central RNA polymerase II termination factors. Our study uncovers transcription termination as a developmentally regulated process required for cell type-specific gene expression.
2024-04-13 | PXD049375 | Pride
Project description:The Drosophila spermatogenesis cell differentiation pathway involves the activation of a large set of genes in primary spermatocytes. Most of these genes are activated by testis-specific TATA-binding protein associated factors (tTAFs). In the current model for the activation mechanism, Polycomb plays a key role silencing these genes in the germline precursors, and tTAF-dependent activation in primary spermatocytes involves the displacement of Polycomb from gene promoters. We investigated the genome-wide binding of Polycomb in wild type and tTAF mutant testes. According to the model we expected to see a clear enhancement in Polycomb binding at tTAF-dependent spermatogenesis genes in tTAF mutant testes. However, we find little evidence for such an enhancement in tTAF mutant testes compared to wild type. To avoid problems arising from cellular heterogeneity in whole testis analysis, we further tested the model by analysing Polycomb binding in purified germline precursors, representing cells before tTAF-dependent gene activation. Although we find Polycomb associated with its canonical targets, we find little or no evidence of Polycomb at spermatogenesis genes. The lack of Polycomb at tTAF-dependent spermatogenesis genes in precursor cells argues against a model where Polycomb displacement is the mechanism of spermatogenesis gene activation.
2013-10-18 | GSE39935 | GEO
Project description:The Drosophila spermatogenesis cell differentiation pathway involves the activation of a large set of genes in primary spermatocytes. Most of these genes are activated by testis-specific TATA-binding protein associated factors (tTAFs). In the current model for the activation mechanism, Polycomb plays a key role silencing these genes in the germline precursors, and tTAF-dependent activation in primary spermatocytes involves the displacement of Polycomb from gene promoters. We investigated the genome-wide binding of Polycomb in wild type and tTAF mutant testes. According to the model we expected to see a clear enhancement in Polycomb binding at tTAF-dependent spermatogenesis genes in tTAF mutant testes. However, we find little evidence for such an enhancement in tTAF mutant testes compared to wild type. To avoid problems arising from cellular heterogeneity in whole testis analysis, we further tested the model by analysing Polycomb binding in purified germline precursors, representing cells before tTAF-dependent gene activation. Although we find Polycomb associated with its canonical targets, we find little or no evidence of Polycomb at spermatogenesis genes. The lack of Polycomb at tTAF-dependent spermatogenesis genes in precursor cells argues against a model where Polycomb displacement is the mechanism of spermatogenesis gene activation. This genome-wide ChIP-array study investigates the binding of Polycomb in three biological samples: wild type (WT) whole testes, tTAF (can) mutant whole testes, and FACS-sorted germline precursor cells. We performed two biological replicates for each sample, except wild type whole testes where we performed three. For all ChIP-array experiments, input chromatin was used as the reference control to assay ChIP enrichment. We used Cy3/Cy5-labelled ChIP and input DNA for hybridisation onto Nimblegen arrays, and we performed a Cy3/Cy5 dye swap for one biological replicate of each sample (see supplementary file: GSE39935_README.txt).
2013-10-18 | E-GEOD-39935 | biostudies-arrayexpress
Project description:Animal germline development and fertility rely on paralogs of general transcription factors that recruit RNA polymerase II to ensure cell type-specific gene expression. It remains unclear whether gene expression processes downstream of such paralog-based transcription is distinct from that of canonical RNA polymerase II genes. In Drosophila, the testis-specific TBP-associated factors (tTAFs) activate over a thousand spermatocyte-specific gene promoters to enable meiosis and germ cell differentiation. Here we show that efficient termination of tTAF-activated transcription requires a testis-specific Polymerase Associated Factor 1 Complex (tPAF) composed of paralogs of canonical Polymerase Associated Factor 1 Complex (PAF1C) proteins. Defective transcription termination in tPAF mutants causes aberrant expression of hundreds of downstream genes due to read-in transcription, compromising cell type-specific gene expression in spermatocytes. Consistently, tPAF is required for the segregation of meiotic chromosomes, sperm individualisation, and male fertility. Comparative in vivo proximity labeling assays of tPAF and PAF1C showed tPAF-specific association with tTAF as well as connections to central RNA polymerase II termination factors. Our study uncovers transcription termination as a developmentally regulated process required for cell type-specific gene expression.
2024-04-16 | GSE263955 | GEO