Functional interaction of human Ssu72 with RNA polymerase II complexes

Benjamin M Spector; Michael E Turek; David H Price

doi:10.1371/journal.pone.0213598

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Functional interaction of human Ssu72 with RNA polymerase II complexes

Journal article

Open access

Peer reviewed

Functional interaction of human Ssu72 with RNA polymerase II complexes

Benjamin M Spector, Michael E Turek and David H Price

PloS one, Vol.14(3), pp.e0213598-e0213598

2019

DOI: 10.1371/journal.pone.0213598

PMCID: PMC6430399

PMID: 30901332

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Abstract

Phosphorylation of the C-terminal domain (CTD) of the large subunit of human RNA polymerase II (Pol II) is regulated during the transcription cycle by the combined action of specific kinases and phosphatases. Pol II enters into the preinitiation complex (PIC) unphosphorylated, but is quickly phosphorylated by Cdk7 during initiation. How phosphatases alter the pattern and extent of CTD phosphorylation at this early stage of transcription is not clear. We previously demonstrated the functional association of an early-acting, magnesium-independent phosphatase with early elongation complexes. Here we show that Ssu72 is responsible for that activity. We found that the phosphatase enters the transcription cycle during the formation of PICs and that Ssu72 is physically associated with very early elongation complexes. The association of Ssu72 with elongation complexes was stable to extensive washing with up to 200 mM KCl. Interestingly, Ssu72 ceased to function on complexes that contained RNA longer than 28 nt. However, when PICs were washed before initiation, the strict cutoff at 28 nt was lost. This suggests that factor(s) are important for the specific regulation of Ssu72 function during the transition between initiation and pausing. Overall, our results demonstrate when Ssu72 can act on early transcription complexes and suggest that Ssu72 may also function in the PIC prior to initiation.

Carrier Proteins - chemistry

Carrier Proteins - metabolism

Cyclin-Dependent Kinases - chemistry

Cyclin-Dependent Kinases - metabolism

Humans

Phosphorylation

Potassium Chloride - chemistry

Protein Domains

RNA Polymerase II - chemistry

RNA Polymerase II - metabolism

Transcription Elongation, Genetic

Transcription Initiation, Genetic

Details

Title: Subtitle: Functional interaction of human Ssu72 with RNA polymerase II complexes
Creators: Benjamin M Spector - University of Iowa
Michael E Turek - University of Iowa
David H Price - University of Iowa
Resource Type: Journal article
Publication Details: PloS one, Vol.14(3), pp.e0213598-e0213598
DOI: 10.1371/journal.pone.0213598
PMID: 30901332
PMCID: PMC6430399
NLM abbreviation: PLoS One
ISSN: 1932-6203
eISSN: 1932-6203
Grant note: R35 GM126908 / NIGMS NIH HHS R01 GM035500 / NIGMS NIH HHS
Language: English
Date published: 2019
Academic Unit: Biochemistry and Molecular Biology
Record Identifier: 9984230629902771

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