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RPAP2 regulates a transcription initiation checkpoint by inhibiting assembly of pre-initiation complex

ABSTRACT: RPAP2 regulates a transcription initiation checkpoint by inhibiting assembly of pre-initiation complex

PROVIDER: PRJNA797170 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17382433_1.fastq.gz	Fastqsanger.gz
	SRR17382433_2.fastq.gz	Fastqsanger.gz
	SRR17382434_1.fastq.gz	Fastqsanger.gz
	SRR17382434_2.fastq.gz	Fastqsanger.gz
	SRR17382435_1.fastq.gz	Fastqsanger.gz

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RPAP2 regulates a transcription initiation checkpoint by prohibiting assembly of preinitiation complex [ChIP-Rx]

Project description:RNA polymerase II (Pol II)-mediated transcription in metazoan requires precise regulation. RNA polymerase II-associated protein 2 (RPAP2) was previously identified to transport Pol II from cytoplasm to nucleus and dephosphorylates Pol II C-terminal domain (CTD). We found that RPAP2 binds hypo/hyper-phosphorylated Pol II with undetectable phosphatase activity. Structure of RPAP2-Pol II shows mutually exclusive assembly of RPAP2-Pol II and pre-initiation complex (PIC) due to three steric clashes. RPAP2 prevents/disrupts Pol II-TFIIF interaction and impairs in vitro transcription initiation, suggesting a function in prohibiting PIC assembly. Loss of RPAP2 in cells leads to global accumulation of TFIIF and Pol II at promoters, indicating critical role of RPAP2 in inhibiting PIC assembly independent of its putative phosphatase activity. Our study indicates that RPAP2 functions as a gatekeeper to prohibit PIC assembly and transcription initiation and suggests a novel transcription checkpoint.

2022-05-04 | GSE193678 | GEO

RPAP2 regulates a transcription initiation checkpoint by prohibiting assembly of preinitiation complex [ChIP-seq]

2022-05-04 | GSE193679 | GEO

RPAP2 regulates a transcription initiation checkpoint by prohibiting assembly of preinitiation complex [TT-seq]

2022-05-04 | GSE192782 | GEO

RPAP2 regulates a transcription initiation checkpoint by inhibiting assembly of pre-initiation complex

Project description:This SuperSeries is composed of the SubSeries listed below.

2022-05-03 | GSE193680 | GEO

Dephosphorylation of the pre-initiation complex during S-phase is critical for origin firing dynamics

Project description:Genome stability requires complete DNA duplication exactly once before division. In eukaryotes, cyclin-dependent kinase (CDK) plays a dual role in this regulation by inhibiting helicase loading factors and activating origin firing. CDK drives initiation by phosphorylation of two substrates, Sld2 and Sld3, forming the transient and limiting intermediate called the pre-initiation complex (pre-IC). The importance and mechanism of dissociation of the pre-IC from origins is not understood. Here we show in the budding yeast Saccharomyces cerevisiae that CDK phosphorylations of Sld3 and Sld2 are specifically and rapidly turned-over during interphase by the PP2A and PP4 phosphatases. Inhibition of dephosphorylation of Sld3/Sld2 causes dramatic defects in replication initiation genome-wide, retention of the pre-IC at origins and cell death. These studies not only provide a mechanism to ensure that Sld3/Sld2 are dephosphorylated before helicase loading factors but also establish a novel positive role for phosphatases in eukaryotic origin firing.

2021-11-01 | GSE186490 | GEO

RPAP2 regulates a transcription initiation checkpoint by prohibiting assembly of preinitiation complex [ChIP-Rx]

Project description:RPAP2 regulates a transcription initiation checkpoint by prohibiting assembly of preinitiation complex [ChIP-Rx]

| PRJNA797176 | ENA

RPAP2 regulates a transcription initiation checkpoint by prohibiting assembly of preinitiation complex [ChIP-seq]

Project description:RPAP2 regulates a transcription initiation checkpoint by prohibiting assembly of preinitiation complex [ChIP-seq]

| PRJNA797177 | ENA

RPAP2 regulates a transcription initiation checkpoint by prohibiting assembly of preinitiation complex [TT-seq]

Project description:RPAP2 regulates a transcription initiation checkpoint by prohibiting assembly of preinitiation complex [TT-seq]

| PRJNA793177 | ENA

Structural inventory of native ribosomal ABCE1-43S pre-initiation complex

Project description:In eukaryotic translation, the termination and recycling phases are linked to subsequent initiation by persistence of several factors. These comprise the large eIF3 complex, eIF3j (Hcr1 in yeast) and the ATP-binding cassette protein ABCE1 (Rli1 in yeast). The ATPase is mainly active as a recycling factor, but it can remain bound to the dissociated 40S subunit until formation of 43S pre-initiation complexes. However, its functional role and native architectural context remains largely enigmatic. Here, we present an architectural inventory of native yeast and human ABCE1-containing pre-initiation complexes by cryo-EM. We found that ABCE1 was mostly associated with early 43S but also later 48S phases of initiation. It adopted a novel hybrid conformation of its nucleotide binding sites, which was stabilized by the N-terminus of eIF3j. Further, eIF3j occupied the mRNA entry channel via its ultimate C-terminus explaining its antagonistic role with respect to mRNA binding. Moreover, the native human samples provided a near-complete molecular picture of the architecture and sophisticated interaction network of the 43S-bound eIF3 complex and also the eIF2 ternary complex containing the initiator tRNA.

2020-10-16 | PXD020849 | Pride

Structure of a Complete Mediator-RNA Polymerase II Pre-Initiation Complex, and the Regulation of Transcription

Project description:MedPIC: Crosslinking-MS of 54 subunit pre-initiation complex from yeast. MedPolII: Crosslinking-MS of 33 subunit Mediator + polII complex from yeast.

2016-08-02 | MSV000080013 | MassIVE

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