RNA Polymerase III Transcription from the Human U6 and Adenovirus Type 2 VAI Promoters Has Different Requirements for Human BRF, a Subunit of Human TFIIIB
Copyright policy )RNA Polymerase III Transcription from the Human U6 and Adenovirus Type 2 VAI Promoters Has Different Requirements for Human BRF, a Subunit of Human TFIIIB
Mammalian TFIIIB can be separated into two fractions required for transcription of the adenovirus type 2 VAI gene, which have been designated 0.38M-TFIIIB and 0.48M-TFIIIB. While 0.48M-TFIIIB has not been characterized, 0.38M-TFIIIB corresponds to a TBP-containing complex. We describe here the purification of this complex, which consists of TBP and a closely associated polypeptide of 88 kDa, and the isolation of a cDNA corresponding to the 88-kDa polypeptide. The predicted protein sequence reveals that the 88-kDa polypeptide corresponds to a human homolog of the Saccharomyces cerevisiae BRF protein, a subunit of yeast TFIIIB. Human BRF (hBRF) probably corresponds to TFIIIB90, a protein previously cloned by Wang and Roeder (Proc. Natl. Acad. Sci. USA 92:7026-7030, 1995), although its predicted amino acid sequence differs from that reported for TFIIIB90 over a stretch of 67 amino acids as a result of frameshifts. Immunodepletion of more than 90 to 95% of the hBRF present in a transcription extract severely debilitates transcription from the tRNA-type VAI promoter but does not affect transcription from the TATA box-containing human U6 promoter, suggesting that the 0.38M-TFIIIB complex, and perhaps hBRF as well, is not required for U6 transcription.
- Howard Hughes Medical Institute United States
- Cold Spring Harbor Laboratory United States
TATA-Binding Protein Associated Factors, Saccharomyces cerevisiae Proteins, Base Sequence, Genes, Viral, Transcription, Genetic, Ribonucleoprotein, U4-U6 Small Nuclear, Adenoviruses, Human, Molecular Sequence Data, RNA Polymerase III, Gene Expression Regulation, Transcription Factor TFIIIB, Humans, RNA, Viral, Amino Acid Sequence, Cloning, Molecular, Promoter Regions, Genetic, Sequence Alignment, Transcription Factors
TATA-Binding Protein Associated Factors, Saccharomyces cerevisiae Proteins, Base Sequence, Genes, Viral, Transcription, Genetic, Ribonucleoprotein, U4-U6 Small Nuclear, Adenoviruses, Human, Molecular Sequence Data, RNA Polymerase III, Gene Expression Regulation, Transcription Factor TFIIIB, Humans, RNA, Viral, Amino Acid Sequence, Cloning, Molecular, Promoter Regions, Genetic, Sequence Alignment, Transcription Factors
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