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

Mital, R., Kobayashi, R., Hernandez, N. (December 1996) 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. Molecular and Cellular Biology, 16 (12). pp. 7031-42. ISSN 0270-7306

URL: http://www.ncbi.nlm.nih.gov/pubmed/8943358

Abstract

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.

Item Type: Paper
Uncontrolled Keywords: Adenoviruses, Human/genetics Amino Acid Sequence Base Sequence Cloning, Molecular Gene Expression Regulation Genes, Viral Humans Molecular Sequence Data Promoter Regions (Genetics)/genetics RNA Polymerase III/ genetics RNA, Viral/ genetics Research Support, U.S. Gov't, P.H.S. Ribonucleoprotein, U4-U6 Small Nuclear/ genetics Sequence Alignment TATA-Binding Protein Associated Factors Transcription Factor TFIIIB Transcription Factors/ genetics/isolation & purification Transcription, Genetic
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > transcription
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > RNA polymerase
organism description > yeast > Saccharomyces
organism description > virus > adenovirus
Investigative techniques and equipment > cloning
Investigative techniques and equipment > assays > cloning

bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene expression
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > nuclear ribonucleoprotein
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > transcription factor
CSHL Authors:
Communities: CSHL labs > Hernandez lab
CSHL labs > Kobayashi lab
Depositing User: Kathleen Darby
Date: December 1996
Date Deposited: 13 May 2014 17:05
Last Modified: 13 May 2014 17:05
PMCID: PMC231706
Related URLs:
URI: http://repository.cshl.edu/id/eprint/30091

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