THZ1 Reveals Roles for Cdk7 in Co-transcriptional Capping and Pausing

Kyle A Nilson; Jiannan Guo; Michael E Turek; John E Brogie; Elizabeth Delaney; Donal S Luse; David H Price

doi:10.1016/j.molcel.2015.06.032

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THZ1 Reveals Roles for Cdk7 in Co-transcriptional Capping and Pausing

Journal article

Open access

Peer reviewed

THZ1 Reveals Roles for Cdk7 in Co-transcriptional Capping and Pausing

Kyle A Nilson, Jiannan Guo, Michael E Turek, John E Brogie, Elizabeth Delaney, Donal S Luse and David H Price

Molecular cell, Vol.59(4), pp.576-587

08/20/2015

DOI: 10.1016/j.molcel.2015.06.032

PMCID: PMC4546572

PMID: 26257281

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Abstract

The Cdk7 subunit of TFIIH phosphorylates RNA polymerase II (Pol II) during initiation, and, while recent studies show that inhibition of human Cdk7 negatively influences transcription, the mechanisms involved are unclear. Using in vitro transcription with nuclear extract, we demonstrate that THZ1, a covalent Cdk7 inhibitor, causes defects in Pol II phosphorylation, co-transcriptional capping, promoter proximal pausing, and productive elongation. THZ1 does not affect initiation but blocks essentially all Pol II large subunit C-terminal domain (CTD) phosphorylation. We found that guanylylation of nascent RNAs is length dependent and modulated by a THZ1-sensitive factor present in nuclear extract. THZ1 impacts pausing through a capping-independent block of DSIF and NELF loading. The P-TEFb-dependent transition into productive elongation was also inhibited by THZ1, likely due to loss of DSIF. Capping and pausing were also reduced in THZ1-treated cells. Our results provide mechanistic insights into THZ1 action and how Cdk7 broadly influences transcription and capping.

Phosphorylation

Cyclin-Dependent Kinases - metabolism

Transcription Factors - chemistry

Humans

Transcriptional Elongation Factors - chemistry

Pyrimidines - chemistry

Antineoplastic Agents - pharmacology

Cyclin-Dependent Kinases - antagonists & inhibitors

Transcription Initiation, Genetic

RNA Polymerase II - chemistry

Protein Structure, Tertiary

Promoter Regions, Genetic

Cyclin-Dependent Kinases - chemistry

RNA Processing, Post-Transcriptional

Nuclear Proteins - metabolism

Pyrimidines - pharmacology

Antineoplastic Agents - chemistry

Nuclear Proteins - chemistry

Transcription Factors - metabolism

Transcriptional Elongation Factors - metabolism

Phenylenediamines - pharmacology

Protein Processing, Post-Translational

HeLa Cells

Kinetics

Phenylenediamines - chemistry

Details

Title: Subtitle: THZ1 Reveals Roles for Cdk7 in Co-transcriptional Capping and Pausing
Creators: Kyle A Nilson - Molecular and Cellular Biology Program, University of Iowa, Iowa City, IA 52242, USA
Jiannan Guo - Biochemistry Department, University of Iowa, Iowa City, IA 52242, USA
Michael E Turek - Biochemistry Department, University of Iowa, Iowa City, IA 52242, USA
John E Brogie - Biochemistry Department, University of Iowa, Iowa City, IA 52242, USA
Elizabeth Delaney - Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA
Donal S Luse - Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA
David H Price - Molecular and Cellular Biology Program, University of Iowa, Iowa City, IA 52242, USA; Biochemistry Department, University of Iowa, Iowa City, IA 52242, USA. Electronic address: david-price@uiowa.edu
Resource Type: Journal article
Publication Details: Molecular cell, Vol.59(4), pp.576-587
DOI: 10.1016/j.molcel.2015.06.032
PMID: 26257281
PMCID: PMC4546572
NLM abbreviation: Mol Cell
ISSN: 1097-2765
eISSN: 1097-4164
Publisher: United States
Grant note: R01 GM35500-29 / NIGMS NIH HHS R01 GM035500 / NIGMS NIH HHS
Language: English
Date published: 08/20/2015
Academic Unit: Biochemistry and Molecular Biology
Record Identifier: 9984025259502771

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