RNA polymerase III transcription repressed by Rb through its interactions with TFIIIB and TFIIIC2.
The retinoblastoma susceptibility gene product (Rb) generally represses RNA polymerase III (Pol III)-directed transcription. This implies that Rb interacts with essential transcription factors. Mutations in either the A or B subdomains in the Rb pocket interfere with Rb-mediated repression of Pol III-directed transcription, which indicates that both subdomains are directly involved in this activity. Addition of either purified TFIIIB or purified TFIIIC2 partially relieves Rb-mediated repression and restores activity to nuclear extracts that had been depleted of essential factors by binding to Rb. Pull down and coimmunoprecipitation experiments as well as functional assays indicate that Rb interacts with both TFIIIB and TFIIIC2 and that the A subdomain is primarily required for binding TFIIIB and the B subdomain for binding TFIIIC2. While Rb interacts with both factors, the A subdomain is more important than the B subdomain in directing Rb-mediated repression, and TFIIIB is the principal target of that activity.
Chu
W M
WM
Section of Molecular and Cellular Biology, University of California, Davis, Davis, California 95616, USA.
Wang
Z
Z
Roeder
R G
RG
Schmid
C W
CW
D000595:Amino Acid Sequence; D001665:Binding Sites; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D005982:Glutathione Transferase; D006367:HeLa Cells; D006801:Humans; D007668:Kidney; D007700:Kinetics; D008962:Models, Theoretical; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D012320:RNA Polymerase III / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000378:metabolism*; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors / Q000302:isolation & purification / Q000378:metabolism*; D019474:Transcription Factors, TFIII; D014158:Transcription, Genetic; D014162:Transfection; D000098832:Transcription Factor TFIIIC
D011993:Recombinant Fusion Proteins; D016160:Retinoblastoma Protein; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors; D019474:Transcription Factors, TFIII; D000098832:Transcription Factor TFIIIC; D005982:Glutathione Transferase; D012320:RNA Polymerase III