DNA binding provides a signal for phosphorylation of the RNA polymerase II heptapeptide repeats.
Isolated transcription complexes contain a protein kinase that phosphorylates the heptapeptide repeats of the carboxy-terminal domain (CTD) of the RNA polymerase II (RNAP II) large subunit in an apparently promoter-dependent manner. We now show that the essential features of this reaction can be reproduced in a reconstituted system containing three macromolecular components: a fusion protein consisting of the CTD of RNAP II fused to a heterologous DNA-binding domain, an activating DNA fragment containing the recognition sequence for the fusion protein, and a protein kinase that binds nonspecifically to DNA. This kinase closely resembles a previously known DNA-dependent protein kinase. Evidently, the association of the CTD with DNA provides a key signal for phosphorylation. There appears to be no absolute requirement for specific contacts with other DNA-bound transcription factors.
Peterson
S R
SR
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309.
Dvir
A
A
Anderson
C W
CW
Dynan
W S
WS
D000818:Animals; D001345:Autoradiography; D004247:DNA / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010766:Phosphorylation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011494:Protein Kinases / Q000378:metabolism; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D013698:Templates, Genetic; D014158:Transcription, Genetic
D011993:Recombinant Fusion Proteins; D004247:DNA; D011494:Protein Kinases; D012319:RNA Polymerase II