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
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: |
https://repository.cshl.edu/id/eprint/30091 |
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