Recruitment of P-TEFb (Cdk9-Pch1) to chromatin by the cap-methyl transferase Pcm1 in fission yeast
- 1 March 2007
- journal article
- research article
- Published by Springer Science and Business Media LLC in The EMBO Journal
- Vol. 26 (6), 1552-1559
- https://doi.org/10.1038/sj.emboj.7601627
Abstract
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.Keywords
This publication has 52 references indexed in Scilit:
- Gene-specific requirement for P-TEFb activity and RNA polymerase II phosphorylation within the p53 transcriptional programGenes & Development, 2006
- Cyclin-Dependent Kinase 9 (Cdk9) of Fission Yeast Is Activated by the CDK-Activating Kinase Csk1, Overlaps Functionally with the TFIIH-Associated Kinase Mcs6, and Associates with the mRNA Cap Methyltransferase Pcm1 In VivoMolecular and Cellular Biology, 2006
- Transcription Factor and Polymerase Recruitment, Modification, and Movement on dhsp70 In Vivo in the Minutes following Heat ShockMolecular and Cellular Biology, 2003
- Bur1 Kinase Is Required for Efficient Transcription Elongation by RNA Polymerase IIMolecular and Cellular Biology, 2003
- Interactions between Fission Yeast Cdk9, Its Cyclin Partner Pch1, and mRNA Capping Enzyme Pct1 Suggest an Elongation Checkpoint for mRNA Quality ControlOnline Journal of Public Health Informatics, 2003
- Phosphorylation of the RNA Polymerase II Carboxy-Terminal Domain by the Bur1 Cyclin-Dependent KinaseMolecular and Cellular Biology, 2001
- RNA CAPPING ENZYME AND DNA-LIGASE - A SUPERFAMILY OF COVALENT NUCLEOTIDYL TRANSFERASESMolecular Microbiology, 1995
- In vivo transcriptional pausing and cap formation on three Drosophila heat shock genes.Proceedings of the National Academy of Sciences of the United States of America, 1993
- TATA box mutations in the Schizosaccharomyces pombe nmt1 promoter affect transcription efficiency but not the transcription start point or thiamine repressibilityGene, 1993
- [56] Molecular genetic analysis of fission yeast Schizosaccharomyces pombeMethods in Enzymology, 1991