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Influence of IWS1 TND-interacting motifs (TIMs) on gene expression patterns [RNA-seq]

ABSTRACT: Influence of IWS1 TND-interacting motifs (TIMs) on gene expression patterns [RNA-seq]

PROVIDER: PRJNA745577 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15100958_1.fastq.gz	Fastqsanger.gz
	SRR15100958_2.fastq.gz	Fastqsanger.gz
	SRR15100959_1.fastq.gz	Fastqsanger.gz
	SRR15100959_2.fastq.gz	Fastqsanger.gz
	SRR15100960_1.fastq.gz	Fastqsanger.gz

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Influence of IWS1 TND-interacting motifs (TIMs) on gene expression patterns [RNA-seq]

Project description:To reveal the influence of TND-interacting motifs (TIMs), which bind TFIIS N-terminal domains (TNDs), we prepared KELLY cells expressing either wild-type IWS1-FLAG or IWS1-FLAG containing disruptive mutations in the unstructured TIM3 region. Following lentiviral transduction of either WT or M3 IWS1-FLAG, we prepared mRNA from these samples and performed expression profiling using RNA-seq. Analysis of differential gene expression provided markers that are differentially regulated by TIM3 of IWS1.

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Influence of IWS1 TND-interacting motifs (TIMs) on chromatin localization patterns [ChIP-seq]

Project description:To reveal the influence of TND-interacting motifs (TIMs), which bind TFIIS N-terminal domains (TNDs), we prepared KELLY cells expressing either wild-type IWS1-FLAG or IWS1-FLAG containing disruptive mutations in the unstructured TIM3 region. Following lentiviral transduction of either WT or M3 IWS1-FLAG, we performed ChIP using anti-FLAG antibodies from these samples and performed chromatin occupancy profiling using ChIP-seq. Analysis of locations reveal that both WT and M3 samples are highly enriched in at transcribed gene bodies.

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Influence of IWS1 TND-interacting motifs (TIMs) on transcription elongation [PRO-seq]

Project description:To reveal the influence of TND-interacting motifs (TIMs), which bind TFIIS N-terminal domains (TNDs), we prepared KELLY cells expressing either wild-type IWS1-FLAG or IWS1-FLAG containing disruptive mutations in the unstructured TIM3 region. Following lentiviral transduction of either WT or M3 IWS1-FLAG, we prepared PRO-seq libraries from these samples and examined elongation profiles. Analysis of elongation profiles revealed that TIM3 disruption induces pervasive defects of transcription elongation.

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Project description:Influence of IWS1 TND-interacting motifs (TIMs) on chromatin localization patterns [ChIP-seq]

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Project description:Influence of IWS1 TND-interacting motifs (TIMs) on transcription elongation [PRO-seq]

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Chromatin and transcriptional mechanisms regulated by TFIIS N-terminal domains (TNDs) and TND-interacting motifs (TIMs)

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IWS1 Stimulates Pol II Transcription Elongation In Vivo

Project description:The protein IWS1 (Interacts with SPT6 1) has been implicated in various transcription-associated processes, but whether it has a direct role in RNA polymerase (Pol) II transcription remains unclear. Here, we establish a rapid depletion system for IWS1 in human cells, and use it to elucidate its function in vivo by multi-omics kinetic analysis. We show that IWS1 depletion results in a global decrease of RNA synthesis in vivo that is due to a decrease in Pol II elongation velocity and in vitro biochemical assays reveal that IWS1 directly stimulates Pol II transcription by lowering the nucleosome entry barrier. Further analyses show that H3K36me3 decreases upon IWS1 depletion although the recruitment of the histone methyltransferase SETD2 to chromatin is unaffected. We conclude that IWS1 has a direct role in Pol II transcription and is essential for maintaining normal RNA elongation, which in turn is required for normal chromatin methylation.

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Project description:Interacts with SPT6 1 (IWS1) has been implicated in various co-transcriptional processes, yet its direct role in RNA Polymerase (Pol) II transcription in vivo remains elusive. In this study, we utilized a rapid depletion system of IWS1 in human cells, followed by multi-omics analysis and biochemical assays, to elucidate its function. We show that IWS1 depletion results in a global decrease of RNA synthesis, but does not affect the chromatin recruitment of the histone methyltransferase SETD2. In vivo and in vitro analyses of nascent transcription and histone modifications revealed that the observed decrease in methylation is a secondary effect of the transcription defect caused by IWS1 depletion. Whereas these results show that IWS1 is essential for maintaining normal RNA Pol II elongation velocity in vivo, and that it can directly stimulate transcription.

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