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ZWC Complex-Mediated SPT5 Phosphorylation Suppresses Divergent Antisense RNA Transcription at the Active Gene Promoters [RNA-Seq]

ABSTRACT: Purpose: The goals of this study are to understand the mechanistic roles of newly identified ZWC chromatin complex on the regulation of transcriptional process in actively transcribed genes. The novel ZWC complex is composed of ZC3H4, WDR82, and casein kinase II (CK2) subunits. Methods: Three biological replicates of RNA-seq data were generated after HEK293 cells were treated with control or ZC3H4 siRNAs. The RNA-Seq libraries were generated from 200 ng of total RNA using TruSeq Stranded Total RNA Library Prep kit (Illumina) according to the manufacturer's protocol. Results: The ZWC complex preferentially localizes at TSSs of active genes through the direct interactions of ZC3H4 and WDR82 subunits with S5p RNAPII CTD. ZC3H4 depletion leads to increased divergent antisense transcription. Conclusion: Our study provides evidence that the newly identified ZWC-DSIF axis regulates the direction of transcription during the transition from early to productive elongation

ORGANISM(S): Homo sapiens

PROVIDER: GSE186808 | GEO | 2022/03/28

REPOSITORIES: GEO

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ZWC Complex-Mediated SPT5 Phosphorylation Suppresses Divergent Antisense RNA Transcription at the Active Gene Promoters [ChIP-Seq]

Project description:Purpose: The goals of this study are to understand the mechanistic roles of newly identified ZWC chromatin complex on the regulation of transcriptional process in actively transcribed genes. The novel ZWC complex is composed of ZC3H4, WDR82, and casein kinase II (CK2) subunits. Methods: Two biological replicates of ChIP-seq data for ZC3H4, WDR82, CK2, and SPT5 were generated from HEK293 cells. And one replicate of ChIP-seq data for RNAPII, H3K4me3, H3K27me3, and H3K36me3 was generated in HEK293 cells. Two biological replicates of ChIP-seq data were generated for RNAPII, S5p RNAPII, S2p RNAPII, SPT5, H3K4me3, H3K36me3, H3K27me3 after HEK293 cells were treated with control or ZC3H4 siRNAs. Results: The ZWC complex preferentially localizes at TSSs of active genes through the direct interactions of ZC3H4 and WDR82 subunits with S5p RNAPII CTD. ZC3H4 depletion leads to increased divergent antisense transcription. Conclusion: Our study provides evidence that the newly identified ZWC-DSIF axis regulates the direction of transcription during the transition from early to productive elongation

2022-03-28 | GSE186758 | GEO

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ZC3H4, ARS2, and WDR82 coordinate non-coding transcriptional termination and are opposed by telescripting on protein-coding genes [chip-seq]

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ZC3H4, ARS2, and WDR82 coordinate non-coding transcriptional termination and are opposed by telescripting on protein-coding genes [ARS2_POINT-Seq]

Project description:There are thousands of unstable non-coding (nc) RNAs in humans, but their transcriptional regulation is poorly characterized compared to protein-coding transcripts. We recently identified a restrictor complex containing ZC3H4 and WDR82, which terminates non-coding transcription. Here we show that the ncRNA termination factor, ARS2, and the nuclear exosome targeting complex (NEXT) are part of this complex.ARS2/NEXT and WDR82 contact different parts of ZC3H4, which are required for its transcription regulatory function. They facilitateZC3H4-mediated termination by different mechanisms: ARS2 aids substrate targeting and WDR82 mediates RNA polymerase II interaction.ZC3H4 and WDR82 predominantly affect antisense, unstable, and intragenic ncRNAs. We provide an explanation for this by showing that U1 telescripting shields hundreds of protein-coding transcripts from their termination functions.We have defined an expanded ZC3H4 restrictor complex, its principles of action, and how protein-coding transcripts evade it.

2023-07-05 | GSE228286 | GEO

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Project description:There are thousands of unstable non-coding (nc) RNAs in humans, but their transcriptional regulation is poorly characterized compared to protein-coding transcripts. We recently identified a restrictor complex containing ZC3H4 and WDR82, which terminates non-coding transcription. Here we show that the ncRNA termination factor, ARS2, and the nuclear exosome targeting complex (NEXT) are part of this complex.ARS2/NEXT and WDR82 contact different parts of ZC3H4, which are required for its transcription regulatory function. They facilitateZC3H4-mediated termination by different mechanisms: ARS2 aids substrate targeting and WDR82 mediates RNA polymerase II interaction.ZC3H4 and WDR82 predominantly affect antisense, unstable, and intragenic ncRNAs. We provide an explanation for this by showing that U1 telescripting shields hundreds of protein-coding transcripts from their termination functions.We have defined an expanded ZC3H4 restrictor complex, its principles of action, and how protein-coding transcripts evade it.

2023-07-05 | GSE228293 | GEO

ZC3H4, ARS2, and WDR82 coordinate non-coding transcriptional termination and are opposed by telescripting on protein-coding genes [PNUTS_U1_AMO]

Project description:There are thousands of unstable non-coding (nc) RNAs in humans, but their transcriptional regulation is poorly characterized compared to protein-coding transcripts. We recently identified a restrictor complex containing ZC3H4 and WDR82, which terminates non-coding transcription. Here we show that the ncRNA termination factor, ARS2, and the nuclear exosome targeting complex (NEXT) are part of this complex.ARS2/NEXT and WDR82 contact different parts of ZC3H4, which are required for its transcription regulatory function. They facilitateZC3H4-mediated termination by different mechanisms: ARS2 aids substrate targeting and WDR82 mediates RNA polymerase II interaction.ZC3H4 and WDR82 predominantly affect antisense, unstable, and intragenic ncRNAs. We provide an explanation for this by showing that U1 telescripting shields hundreds of protein-coding transcripts from their termination functions.We have defined an expanded ZC3H4 restrictor complex, its principles of action, and how protein-coding transcripts evade it.

2023-07-05 | GSE228291 | GEO

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Project description:There are thousands of unstable non-coding (nc) RNAs in humans, but their transcriptional regulation is poorly characterized compared to protein-coding transcripts. We recently identified a restrictor complex containing ZC3H4 and WDR82, which terminates non-coding transcription. Here we show that the ncRNA termination factor, ARS2, and the nuclear exosome targeting complex (NEXT) are part of this complex.ARS2/NEXT and WDR82 contact different parts of ZC3H4, which are required for its transcription regulatory function. They facilitateZC3H4-mediated termination by different mechanisms: ARS2 aids substrate targeting and WDR82 mediates RNA polymerase II interaction.ZC3H4 and WDR82 predominantly affect antisense, unstable, and intragenic ncRNAs. We provide an explanation for this by showing that U1 telescripting shields hundreds of protein-coding transcripts from their termination functions.We have defined an expanded ZC3H4 restrictor complex, its principles of action, and how protein-coding transcripts evade it.

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