A minimal RNA polymerase III transcription system from human cells reveals positive and negative regulatory roles for CK2.
In higher eukaryotes, RNA polymerase (pol) III is known to use different transcription factors to recognize three basic types of promoters, but in no case have these transcription factors been completely defined. We show that a highly purified pol III complex combined with the recombinant transcription factors SNAP(c), TBP, Brf2, and Bdp1 directs multiple rounds of transcription initiation and termination from the human U6 promoter. The pol III complex contains traces of CK2, and CK2 associates with the U6 promoter region in vivo. Transcription requires CK2 phosphorylation of the pol III complex. In contrast, CK2 phosphorylation of TBP, Brf2, and Bdp1 combined is inhibitory. The results define a minimum core machinery, the ultimate target of regulatory mechanisms, capable of directing all steps of the transcription process-initiation, elongation, and termination-by a metazoan RNA polymerase, and suggest positive and negative regulatory roles for CK2 in transcription by pol III.
Hu
Ping
P
Howard Hughes Medical Institute, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA .
Wu
Si
S
Hernandez
Nouria
N
D000074183:ATPases Associated with Diverse Cellular Activities; D000818:Animals; D047390:Casein Kinase II; D002474:Cell-Free System; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005057:Eukaryotic Cells / Q000201:enzymology*; D025461:Feedback, Physiological / Q000235:genetics; D005809:Genes, Regulator / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000235:genetics*; D046988:Proteasome Endopeptidase Complex; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D012320:RNA Polymerase III / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D035361:Transcription Factor TFIIIB / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000235:genetics*
C474805:Bdp1 protein, S cerevisiae; D004268:DNA-Binding Proteins; C106703:SNAPC protein complex, human; D029701:Saccharomyces cerevisiae Proteins; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors; C091750:ZFP36L2 protein, human; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D012320:RNA Polymerase III; D046988:Proteasome Endopeptidase Complex; D000074183:ATPases Associated with Diverse Cellular Activities; C064757:PSMC3 protein, human