Three RNA polymerase II carboxyl-terminal domain kinases display distinct substrate preferences Journal Article
| Authors: | Ramanathan, Y.; Rajpara, S. M.; Reza, S. M.; Lees, E.; Shuman, S.; Mathews, M. B.; Pe'ery, T. |
| Article Title: | Three RNA polymerase II carboxyl-terminal domain kinases display distinct substrate preferences |
| Abstract: | CDK7, CDK8, and CDK9 are cyclin-dependent kinases (CDKs) that phosphorylate the C-terminal domain (CTD) of RNA polymerase II. They have distinct functions in transcription. Because the three CDKs target only serine 5 in the heptad repeat of model CTD substrates containing various numbers of repeats, we tested the hypothesis that the kinases differ in their ability to phosphorylate CTD heptad arrays. Our data show that the kinases display different preferences for phosphorylating individual heptads in a synthetic CTD substrate containing three heptamer repeats and specific regions of the CTD in glutathione S-transferase fusion proteins. They also exhibit differences in their ability to phosphorylate a synthetic CTD peptide that contains Ser-2-PO4. This phosphorylated peptide is a poor substrate for CDK9 complexes. CDK8 and CDK9 complexes, bound to viral activators E1A and Tat, respectively, target only serine 5 for phosphorylation in the CTD peptides, and binding to the viral activators does not change the substrate preference of these kinases. These results imply that the display of different CTD heptads during transcription, as well as their phosphorylation state, can affect their phosphorylation by the different transcription-associated CDKs. |
| Keywords: | controlled study; protein phosphorylation; human cell; genetics; molecular genetics; protein domain; metabolism; cyclin dependent kinase 8; cyclin dependent kinase 9; carboxy terminal sequence; genetic transcription; transcription, genetic; enzyme substrate; hela cell; hela cells; protein serine threonine kinase; phosphorylation; rna; amino acid sequence; molecular sequence data; protein-serine-threonine kinases; transactivator protein; substrate specificity; glutathione transferase; enzyme specificity; cyclin-dependent kinases; cyclin dependent kinase; enzyme substrate complex; biochemistry; rna polymerase ii; enzymes; carboxylic acids; virus protein; cyclin dependent kinase 7; cyclin dependent kinase activating kinase; cyclin-dependent kinase-activating kinase; cyclin-dependent kinase 9; synthetic peptide; humans; human; priority journal; article; distinct substrate preferences; viral activators; cdk8 protein, human; cdk9 protein, human |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 276 |
| Issue: | 14 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2001-04-06 |
| Start Page: | 10913 |
| End Page: | 10920 |
| Language: | English |
| DOI: | 10.1074/jbc.M010975200 |
| PUBMED: | 11278802 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 21 May 2015 -- Source: Scopus |
