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Genome-wide analysis of SAGA subunits: ada2b, spt3, nonstop, and sgf11, as well as RNA pol II in 1-3 hr embryos of Drosophila melanogaster.

ABSTRACT: The Spt-Ada-Gcn5-acetyltransferase (SAGA) chromatin-modifying complex is a transcriptional coactivator that contains four different modules of subunits. The intact SAGA complex has been well characterized for its function in transcription regulation and development. However, little is known about the roles of individual modules within SAGA and if they have any SAGA independent functions. Here we demonstrate that the two enzymatic modules of Drosophila SAGA are differently required in oogenesis. Loss of the HAT activity blocks oogenesis, while loss of the DUB activity does not. However, the DUB module regulates a subset of genes in early embryogenesis and loss of the DUB subunits causes defects in embryogenesis. ChIP-seq analysis of ada2b, spt3, nonstop, and sgf11revealed that both the DUB and HAT modules bind most SAGA target genes even though many of these targets do not require the DUB module for expression. Furthermore, we found that the DUB module can bind to chromatin and regulate transcription independently of the rest of SAGA. Our results suggest that the DUB module has functions within SAGA as well as independent functions.

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE98862 | GEO | 2017/10/01

SECONDARY ACCESSION(S): PRJNA386532

REPOSITORIES: GEO

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RNA Seq analysis of whole ovaries following mutation of SAGA subunits: Ada2b, Ataxin-7, or non-stop in Drosophila melanogaster

Project description:The Spt-Ada-Gcn5-acetyltransferase (SAGA) chromatin-modifying complex is a transcriptional coactivator that contains four different modules of subunits. The intact SAGA complex has been well characterized for its function in transcription regulation and development. However, little is known about the roles of individual modules within SAGA and if they have any SAGA independent functions. Here we demonstrate that the two enzymatic modules of Drosophila SAGA are differently required in oogenesis. Ada2b is part of the HAT module (Histone Acetyl Transferase), whereas Ataxin-7 and non-stop are members of the DUB module (Deubiquitinase). Loss of the HAT activity blocks oogenesis, while loss of the DUB activity does not. However, the DUB module regulates a subset of genes in early embryogenesis and loss of the DUB subunits causes defects in embryogenesis. ChIP-seq analysis of ada2b, spt3, nonstop, and sgf11revealed that both the DUB and HAT modules bind most SAGA target genes even though many of these targets do not require the DUB module for expression. Furthermore, we found that the DUB module can bind to chromatin and regulate transcription independently of the rest of SAGA. Our results suggest that the DUB module has functions within SAGA as well as independent functions.

2017-10-01 | GSE98864 | GEO

Genome-wide analysis of SAGA subunit ada2b, and histones ubH2B, H2b, H3, H3K9ac, and H3K14ac in wt and Ataxin 7 mutants of 1-3 hr embryos of Drosophila melanogaster

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RNA Seq analysis of 2.5 hour embryos of Drosophila melanogaster in the absence of Ataxin-7 or non-stop.

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SAGA Complex Subunits in Candida albicans Differentially Regulate Filamentation, Invasiveness and Biofilm Formation

Project description:In this study we were interested in studying the role of SAGA subunits in various cellular processes- morphogenetic changes, growth, invasiveness, biofilm formation and to check the role of these subunits under various cellular and genotoxic conditions. In this work, we investigated conditional and null mutants of components of the SAGA complex modules; Ngg1 of the HAT module, Ubp8 of the Dub module, Tra1 of the recruitment module, Spt7 of the architecture module, and Spt8 of the TBP interaction unit to assess their role in processes such as filamentation, invasiveness, and biofilm formation. Spt7 and Spt8 deletion in Candida albicans resulted in ifilamentatation, invasiveness in YPD media at 30 degrees Celsius and also showed sensitivity to antifungal drugs. Wild type SN148 do not make any filaments in YPD at 30 degrees Celsius and didn't show antifungal sensitivity . The aim was to look for transcription profiling of SAGA mutants against wild type to find genes up and down regulated in the mutant especially those ones critical for filamentation, invasiveness and antifungal drug resistance.

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Project description:SAGA and ATAC are two related transcriptional coactivator complexes, sharing the same histone acetyltransferase (HAT) subunit. The HAT activities of SAGA and ATAC are required for metazoan development but the precise role of the two complexes in RNA polymerase II transcription in mammals is less understood. To determine whether SAGA and ATAC have redundant or specific functions dependent on their HAT activities, we compared the effects of HAT inactivation in each complex with that of inactivation of either SAGA or ATAC core subunits in mouse embryonic stem cells (ESCs). We show that core subunits of SAGA or ATAC subunits are required for complex assembly, mouse ESC growth and self-renewal. Additionally, ATAC, but not SAGA subunits are required for ESC viability by regulating the transcription of translation-related genes. Surprisingly, depletion of specific or shared HAT module subunits caused a global decrease in histone H3K9 acetylation, but did not result in significant phenotypic or transcriptional defects. Thus, our results indicate that SAGA and ATAC are differentially required for viability and self-renewal of mouse ESCs by regulating transcription through different pathways, in a HAT-independent manner.

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Project description:SAGA core module subunits regulate gene expression during oogenesis [ChIP-seq]

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