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Distinctive Interactomes of RNA polymerase II phosphorylation during different stages of transcription

ABSTRACT: Distinctive Interactomes of RNA polymerase II phosphorylation during different stages of transcription

PROVIDER: PRJNA913908 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR22812215_1.fastq.gz	Fastqsanger.gz
	SRR22812215_2.fastq.gz	Fastqsanger.gz
	SRR22812216_1.fastq.gz	Fastqsanger.gz
	SRR22812216_2.fastq.gz	Fastqsanger.gz
	SRR22812217_1.fastq.gz	Fastqsanger.gz

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Distinctive Interactomes of RNA polymerase II phosphorylation during different stages of transcription

Project description:Distinctive Interactomes of RNA polymerase II phosphorylation during different stages of transcription

| PRJNA942240 | ENA

Distinctive Interactomes of RNA polymerase II phosphorylation during different stages of transcription

Project description:During eukaryotic transcription, RNA polymerase II undergoes dynamic post-translational modification on the C-terminal domain (CTD) of the largest subunit , generating a sophisticated PTM landscape for the spatiotemporal recruitment to transcriptional regulators. To delineate the protein interactomes recruited to Pol II at different stages of transcription, we in vitro reconstructed phosphorylation patterns of the CTD at Ser5 and Ser2 positions, the hallmark phosphorylation at the initation and productive elongation stages of transcription, respectively. Distinctive protein interactomes indicates different proteins are recruited to RNA polymerase II at different stages of transcription by the phosphorylation of Ser2 and Ser5 of the CTD heptads. Calcium Homeostasis Endoplasmic Reticulum Protein (CHERP) specifically binds to the Ser2 of the heptad. The loss of the interaction between CHERP and Pol II results in broad alternative splicing events. Our method points to a new method to distinguish the PTM codes that coordinate the transcription process.

2023-02-06 | MSV000091213 | MassIVE

Distinctive Interactomes of RNA polymerase II phosphorylation during different stages of transcription

Project description:We have discovered distinctive protein interactomes for Ser2 and Ser5 phosphorylation of the C-terminal domain (CTD) of RNA Pol II. Calcium Homeostasis Endoplasmic reticulum (CHERP) was identified in our pulldown to directly interact with Ser2 phosphomarks on the CTD of Pol II. As Pol II is a chromatin-associated protein, we sought to identify genomic regions that CHERP, a Pol II binding protein, occupy through ChIP-seq analysis.

2023-08-17 | GSE226908 | GEO

Distinctive Interactomes of RNA polymerase II phosphorylation during different stages of transcription

Project description:To investigate the whole-transcriptome effects of Calcium Homeostasis Endoplasmic Reticulum Protein (CHERP)/SCAF6 which specifically binds to the Ser2 of C-terminal domain of RNA Polymerase II, we established HEK293 cell line in which SCAF6 gene was knocked down by shRNA. We then performed gene expression and alternative splicing profiling using data obtained from RNA-seq of 2 cell lines (HEK293 Control vs HEK293 SCAF6 KD).

2023-08-17 | GSE221328 | GEO

Distinctive interactomes of RNA polymerase II phosphorylation during different stages of transcription.

Project description:During eukaryotic transcription, RNA polymerase II undergoes dynamic post-translational modifications on the C-terminal domain (CTD) of the largest subunit, generating an information-rich PTM landscape that transcriptional regulators bind. The phosphorylation of Ser5 and Ser2 of CTD heptad occurs spatiotemporally with the transcriptional stages, recruiting different transcriptional regulators to Pol II. To delineate the protein interactomes at different transcriptional stages, we reconstructed phosphorylation patterns of the CTD at Ser5 and Ser2 in vitro. Our results showed that distinct protein interactomes are recruited to RNA polymerase II at different stages of transcription by the phosphorylation of Ser2 and Ser5 of the CTD heptads. In particular, we characterized calcium homeostasis endoplasmic reticulum protein (CHERP) as a regulator bound by phospho-Ser2 heptad. Pol II association with CHERP recruits an accessory splicing complex whose loss results in broad changes in alternative splicing events. Our results shed light on the PTM-coded recruitment process that coordinates transcription.

| S-EPMC10470302 | biostudies-literature

CDK12 maintains phosphorylation of RNA Polymerase II during epidermal differentiation

Project description:CDK12 plays an important role in regulating epidermal differntaition through maintaining the phosphorylation of RNA polymerase II levels in differentiation associated genes.

2024-02-08 | GSE166407 | GEO

CDK12 maintains phosphorylation of RNA Polymerase II during epidermal differentiation

Project description:CDK12 maintains phosphorylation of RNA Polymerase II during epidermal differentiation

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Transcription Factor Protein Interactomes Reveal Genetic Determinants in Heart Disease

Project description:Congenital Heart Disease (CHD) is present in 1% of live births, and while large-scale genetic studies have uncovered genes associated with CHDs, identifying causal mutations remains a challenge. We hypothesized that genetic determinants for CHDs could be found in the protein interactomes of GATA4 and TBX5, two cardiac transcription factors (TFs) that cause CHDs. Defining their interactomes in human cardiac progenitors via affinity purification-mass spectrometry and integrating the results with genetic data from the Pediatric Cardiac Genomic Consortium (PCGC) revealed an enrichment of de novo variants among proteins that interact with GATA4 or TBX5. A consolidative score designed to prioritize TF interactome members based on distinctive variant, gene, and proband features identified numerous likely CHD-causing genes, including the epigenetic reader GLYR1. GLYR1 and GATA4 widely co-occupied cardiac developmental genes, resulting in co-activation, and the GLYR1 variant associated with CHD disrupted interaction with GATA4. This integrative proteomic and genetic approach provides a framework for prioritizing and interrogating the contribution of genetic variants in CHD and can be extended to other genetic diseases.

2022-02-18 | GSE159411 | GEO

Tyr1 phosphorylation promotes the phosphorylation of Ser2 on the C-terminal domain of RNA polymerase II by P-TEFb

Project description:The Positive Transcription Elongation Factor b (P-TEFb) phosphorylates Ser2 residues of RNA polymerase II’s C-terminal domain (CTD) and is essential for the transition from transcription initiation to elongation in vivo. Surprisingly, P-TEFb exhibits Ser5 phosphorylation activity in vitro. The mechanism garnering Ser2 specificity to P-TEFb remains elusive and hinders understanding the transition from transcription initiation to elongation. Through in vitro reconstruction of CTD phosphorylation, mass spectrometry analysis, and chromatin immunoprecipitation-seq analysis we uncover a mechanism by which Tyr1 phosphorylation directs the kinase activity of P-TEFb and alters its specificity from Ser5 to Ser2. The loss of Tyr1 phosphorylation causes the reduction of Ser2 and accumulation of RNA polymerase II in the promoter region as detected by ChIP-seq. We demonstrate the ability of Tyr1 phosphorylation to generate a heterogeneous CTD modification landscape that expands the CTD’s coding potential. These findings provide direct experimental evidence for a combinatorial CTD phosphorylation code wherein previously installed modifications direct the identity and abundance of subsequent coding events by influencing the behavior of downstream enzymes.

2019-11-01 | GSE131838 | GEO

Sexual differentiation controlled by RNA polymerase II CTD phosphorylation on serine 2

Project description:The switch from cellular proliferation to differentiation occurs to a large extend through specific programs of gene expression. In fission yeast, the high-mobility-group transcription factor Ste11 is the master regulator of sexual differentiation. ste11 is induced by environmental conditions, mostly nitrogen starvation, leading to mating and meiosis 1. We have used ChIP-chip and gene expression profiling to show that CDK-dependent phosphorylation of serine 2 in the C-terminal domain (CTD) of the largest subunit of the RNA polymerase II (PolII) holoenzyme plays a critical role in the induction of ste11 transcription during sexual differentiation while it has a minor impact on gene expression during vegetative growth. Moreover, we demonstrate that both the recruitment of the CTD serine 2 kinase and the phosphorylation event initiate at the promoter region of ste11 in contrast to the classical case where serine 2 phosphorylation occurs across the coding region 2. In the absence of CTD serine 2 phosphorylation, both PolII occupancy at the ste11 locus, and ste11 expression are impaired. This results in sterility that is rescued when ste11 is expressed from the canonical adh promoter. We conclude that a modification of the RNA polymerase II holoenzyme plays a specific and pivotal role in the sexual differentiation. For the ChIP-chip experiments, 3-4 biological replicates were performed for each tagged protein of interest. For the expression experiments, two biological samples were hybridized for each mutant strain (replicates 1 and 2), with two dye-swap technical replicates per sample (replicates 3 and 4).

2010-01-01 | E-GEOD-16498 | biostudies-arrayexpress

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