Recruitment of P-TEFb (Cdk9-Pch1) to chromatin by the cap-methyl transferase Pcm1 in fission yeast

Autor: Martine Raes, Damien Hermand, Marc Dieu, Lionel Tafforeau, Michel Werner, Monique Dewez, Jean Vandenhaute, Allan Guiguen, Julie Soutourina
Rok vydání: 2007
Předmět:
Uracil -- analogs & derivatives
Chromatin Immunoprecipitation
Transcription
Genetic
Positive Transcriptional Elongation Factor B
Blotting
Western
RNA Polymerase II -- metabolism
Cdk9
RNA polymerase II
Article
General Biochemistry
Genetics and Molecular Biology
P-TEFb
Positive Transcriptional Elongation Factor B -- metabolism
Two-Hybrid System Techniques
Schizosaccharomyces
Immunoprecipitation
Elongation
Phosphorylation
Uracil
Methyltransferases -- metabolism
Molecular Biology
Cap-methyltransferase
General Immunology and Microbiology
biology
General Neuroscience
Biologie moléculaire
Methyltransferases
Chromatin -- metabolism
biology.organism_classification
Molecular biology
Chromatin
CTD
Cell biology
Transcription
Genetic -- physiology
biology.protein
Cyclin-dependent kinase 9
RNA Polymerase II
Chromatin immunoprecipitation
Zdroj: EMBO journal, 26 (6
ISSN: 1460-2075
0261-4189
DOI: 10.1038/sj.emboj.7601627
Popis: Capping of nascent pre-mRNAs is thought to be a prerequisite for productive elongation and associated serine 2 phosphorylation of the C-terminal domain (CTD) of RNA polymerase II (PolII). The mechanism mediating this link is unknown, but is likely to include the capping machinery and P-TEPb. We report that the fission yeast P-TEFb (Cdk9-Pch1) forms a complex with the cap-methyltransferase Pcm1 and these proteins colocalise on chromatin. Ablation of Cdk9 function through chemical genetics causes growth arrest and abolishes serine 2 phosphorylation on the PolII CTD. Strikingly, depletion of Pcm1 also leads to a dramatic decrease of phospho-serine 2. Chromatin immunoprecipitations show a severe decrease of chromatin-bound Cdk9-Pch1 when Pcm1 is depleted. On the contrary, Cdk9 is not required for association of Pcm1 with chromatin. Furthermore, compromising Cdk9 activity leads to a promoter-proximal PolII stalling and sensitivity to 6-azauracil, reflecting elongation defects. The in vivo data presented here strongly support the existence of a molecular mechanism where the cap-methyltransferase recruits P-TEFb to chromatin, thereby ensuring that only properly capped transcripts are elongated.
Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
FLWIN
info:eu-repo/semantics/published
Databáze: OpenAIRE
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