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PMID: 7724559 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Human general transcription factor TFIIA: characterization of a cDNA encoding the small subunit and requirement for basal and activated transcription.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 92 ·No. 8 ·1995-04-11 ·Pages 3313-7

DeJong J, Bernstein R, Roeder RG

Abstract

The human general transcription factor TFIIA is one of several factors involved in specific transcription by RNA polymerase II, possibly by regulating the activity of the TATA-binding subunit (TBP) of TFIID. TFIIA purified from HeLa extracts consists of 35-, 19-, and 12-kDa subunits. Here we describe the isolation of a cDNA clone (hTFIIA gamma) encoding the 12-kDa subunit. Using expression constructs derived from hTFIIA gamma and TFIIA alpha/beta (which encodes a 55-kDa precursor to the alpha and beta subunits of natural TFIIA), we have constructed a synthetic TFIIA with a polypeptide composition similar to that of natural TFIIA. The recombinant complex supports the formation of a DNA-TBP-TFIIA complex and mediates both basal and Gal4-VP16-activated transcription by RNA polymerase II in TFIIA-depleted nuclear extracts. In contrast, TFIIA has no effect on tRNA and 5S RNA transcription by RNA polymerase III in this system. We also present evidence that both the p55 and p12 recombinant subunits interact with TBP and that the basic region of TBP is critical for the TFIIA-dependent function of TBP in nuclear extracts.

MeSH Terms
Amino Acid Sequence Base Sequence DNA, Complementary/genetics DNA-Binding Proteins/genetics,metabolism Gene Library Humans Molecular Sequence Data Mutation Protein Binding Recombinant Proteins/metabolism Sequence Homology, Amino Acid TATA Box TATA-Box Binding Protein Transcription Factor TFIIA Transcription Factors/genetics,metabolism Transcription, Genetic
Chemicals
DNA, Complementary DNA-Binding Proteins Recombinant Proteins TATA-Box Binding Protein Transcription Factor TFIIA Transcription Factors
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
DeJong J
Rockefeller University, Laboratory of Biochemistry and Molecular Biology, New York, NY 10021, USA.
Bernstein R
Roeder R G
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.92.8.3313
Published
1995-04-11
Pages
3313-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC42156
Subset
IM
Grants
NCI NIH HHS · CA09637 · United States
NCI NIH HHS · CA42567 · United States
Databases
GENBANK
U21242
External Links
PubMed source DOI: 10.1073/pnas.92.8.3313 PMC Full Text View journal details
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