title: Phosphorylation of the RNA polymerase II C-terminal domain by TFIIH kinase is not essential for transcription of Saccharomyces cerevisiae genome.
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.
authors: Department of Chemistry, Global Research Laboratory for RNAi Medicine, BK21 School of Chemical Materials Science, Sungkyunkwan University, Suwon 440-746, Korea.
authors: Sun Woo
authors: SW
authors: Hong
authors: Seong Min
authors: SM
authors: Hong
authors: Jae Wook
authors: JW
authors: Yoo
authors: Young Chul
authors: YC
authors: Lee
authors: Soyoun
authors: S
authors: Kim
authors: John T
authors: JT
authors: Lis
authors: Dong-Ki
authors: DK
authors: Lee
mesh_terms: D000199:Actins / Q000235:genetics; D001665:Binding Sites; D047369:Chromatin Immunoprecipitation; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005095:Exoribonucleases / Q000235:genetics; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D016681:Genome, Fungal; D051246:Glucose Transport Proteins, Facilitative / Q000235:genetics; D009154:Mutation; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D000098556:mRNA Guanylyltransferases
chemical_list: C465210:Act1 protein, S cerevisiae; D000199:Actins; D004791:Enzyme Inhibitors; D051246:Glucose Transport Proteins, Facilitative; C090594:HXT1 protein, S cerevisiae; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D018844:Cyclin-Dependent Kinases; C084210:Kin28 protein kinase, S cerevisiae; D009713:Nucleotidyltransferases; D012319:RNA Polymerase II; D000098556:mRNA Guanylyltransferases; D005095:Exoribonucleases; C068398:XRN1 protein, S cerevisiae