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PXD047023

PXD047023 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleRepression of pervasive transcription is the primary role of RNA polymerase II CTD serine 2 phosphorylation in fission yeast
DescriptionThe RNA polymerase II carboxy-terminal domain (CTD) consists of conserved repeats of the consensus sequence Y1-S2-P3-T4-S5-P6-S7, which can be phosphorylated to influence distinct stages of the transcription cycle, including RNA processing. Although affinity purification coupled with mass spectrometry has defined CTD-associated proteins, phospho-dependent CTD interactions have remained largely elusive. Proximity-dependent biotinylation (PDB) provides an alternative approach to identify protein-protein associations in the native cellular environment. Here we present a PDB-based map of the fission yeast RNAPII CTD in live cells. The proteomic screen identified known CTD-associated proteins, but also captured new and unexpected CTD proximal proteins. We also used PDB to identify phospho-dependent CTD interactions by using a mutant in which Ser2 was replaced by alanine in every repeat of the fission yeast CTD. Surprisingly, CTD-mediated biotinylation of most 3’ end processing factors was not affected in the S2A mutant, consistent with RNA-seq and ChIP-seq analysis indicating that CTD Ser2 phosphorylation is not required for 3’ end processing and transcription termination. Conversely, we found that CTD Ser2 phosphorylation is critical for the association between RNAPII and the histone methyltransferase Set2 during transcription elongation. We show that loss of CTD Ser2 phosphorylation disables the Set2-Clr6(II) axis, resulting in a global increase in cryptic antisense transcription that correlates with elevated levels of histone acetylation in gene bodies. Our findings reveal that the fundamental role of CTD Ser2 phosphorylation is to establish a chromatin-based repressive state that prevents cryptic intragenic transcription initiation.
HostingRepositoryPRIDE
AnnounceDate2024-10-22
AnnouncementXMLSubmission_2024-10-22_06:38:12.861.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterNouhou EL Moctar Haidara
SpeciesList scientific name: Schizosaccharomyces pombe; NCBI TaxID: NCBITaxon:4896; scientific name: Schizosaccharomyces pombe strain SPY73 975 h+; NCBI TaxID: 881122;
ModificationListacetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
InstrumenttimsTOF Pro; Q Exactive
Dataset History
RevisionDatetimeStatusChangeLog Entry
02023-11-16 07:12:12ID requested
12024-05-06 06:36:32announced
22024-10-22 06:38:13announced2024-10-22: Updated project metadata.
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: TurboID, Set2, antiisense repression, CTD,RNA polymerase II
Contact List
Francois Bachand
contact affiliationRNA Group, Dept of Biochemistry & Functional Genomics, Université de Sherbrooke, Sherbrooke, Québec, Canada
contact emailf.bachand@usherbrooke.ca
lab head
Nouhou EL Moctar Haidara
contact affiliationUniversity of Sherbrooke
contact emailnouhou.el.moctar.haidara@usherbrooke.ca
dataset submitter
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Dataset FTP location
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