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PMID: 19666497 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Phosphorylation of the RNA polymerase II C-terminal domain by TFIIH kinase is not essential for transcription of Saccharomyces cerevisiae genome.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 106 ·No. 34 ·2009-08-25 ·Pages 14276-80

Hong SW, Hong SM, Yoo JW, Lee YC, Kim S, Lis JT, Lee DK

Abstract

Ser-5 phosphorylation of the RNA polymerase II (Pol II) C-terminal domain by TFIIH kinase has been implicated in critical steps in mRNA synthesis, such as Pol II promoter escape and mRNA 5'-capping. However, the general requirement and precise role of TFIIH kinase in Pol II transcription still remain elusive. Here we use a chemical genetics approach to show that, for a majority of budding-yeast genes, specific inhibition of the yeast TFIIH kinase results in a dramatic reduction in both mRNA level and Ser-5 C-terminal domain phosphorylation. Surprisingly, inhibition of TFIIH kinase activity only partially affected both Pol II density and Ser-2 phosphorylation level. The discrepancy between mRNA level and Pol II density is attributed to the defective 5'-capping, which results in the destabilization of mRNAs. Therefore, contrary to the current belief, our study points strongly toward a minor role of TFIIH kinase in Pol II transcription, and a more significant role in mRNA capping in budding yeast.

MeSH Terms
Actins/genetics Binding Sites Chromatin Immunoprecipitation Cyclin-Dependent Kinases/antagonists & inhibitors,genetics,metabolism Enzyme Inhibitors/pharmacology Exoribonucleases/genetics Gene Expression Profiling Gene Expression Regulation, Fungal/drug effects Genome, Fungal Glucose Transport Proteins, Facilitative/genetics Mutation Nucleotidyltransferases/genetics,metabolism Oligonucleotide Array Sequence Analysis Phosphorylation Promoter Regions, Genetic/genetics Protein Binding RNA Polymerase II/genetics,metabolism RNA, Messenger/genetics,metabolism Reverse Transcriptase Polymerase Chain Reaction Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins/antagonists & inhibitors,genetics,metabolism Transcription, Genetic
Chemicals
Act1 protein, S cerevisiae Actins Enzyme Inhibitors Glucose Transport Proteins, Facilitative HXT1 protein, S cerevisiae RNA, Messenger Saccharomyces cerevisiae Proteins Cyclin-Dependent Kinases Kin28 protein kinase, S cerevisiae Nucleotidyltransferases RNA Polymerase II mRNA guanylyltransferase Exoribonucleases XRN1 protein, S cerevisiae
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Hong Sun Woo
Department of Chemistry, Global Research Laboratory for RNAi Medicine, BK21 School of Chemical Materials Science, Sungkyunkwan University, Suwon 440-746, Korea.
Hong Seong Min
Yoo Jae Wook
Lee Young Chul
Kim Soyoun
Lis John T
Lee Dong-Ki
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
1091-6490
DOI
10.1073/pnas.0903642106
Published
2009-08-25
Epub
2009-00-07
Pages
14276-80
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC2732804
Subset
IM
Grants
NIGMS NIH HHS · R01 GM025232 · United States
NIGMS NIH HHS · R01 GM025232-32 · United States
NIGMS NIH HHS · R37 GM025232 · United States
NIGMS NIH HHS · GM25232 · United States
External Links
PubMed source DOI: 10.1073/pnas.0903642106 PMC Full Text View journal details
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