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PMID: 1729606 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The general transcription factor RAP30 binds to RNA polymerase II and prevents it from binding nonspecifically to DNA.

Molecular and cellular biology ·Vol. 12 ·No. 1 ·1992-01-00 ·Pages 30-7

Killeen MT, Greenblatt JF

Abstract

RAP30/74 is a human general transcription factor that binds to RNA polymerase II and is required for initiation of transcription in vitro regardless of whether the promoter has a recognizable TATA box (Z. F. Burton, M. Killeen, M. Sopta, L. G. Ortolan, and J. F. Greenblatt, Mol. Cell. Biol. 8:1602-1613, 1988). Part of the amino acid sequence of RAP30, the small subunit of RAP30/74, has limited homology with part of Escherichia coli sigma 70 (M. Sopta, Z. F. Burton, and J. Greenblatt, Nature (London) 341:410-414, 1989). To determine which sigmalike activities of RAP30/74 could be attributed to RAP30, we purified human RAP30 and a RAP30-glutathione-S-transferase fusion protein that had been produced in E. coli. Bacterially produced RAP30 bound to RNA polymerase II in the absence of RAP74. Both partially purified natural RAP30/74 and recombinant RAP30 prevented RNA polymerase II from binding nonspecifically to DNA. In addition, nonspecific transcription by RNA polymerase II was greatly inhibited by RAP30-glutathione-S-transferase. DNA-bound RNA polymerase II could be removed from DNA by partially purified RAP30/74 but not by bacterially expressed RAP30. Thus, the ability of RAP30/74 to recruit RNA polymerase II to a promoter-bound preinitiation complex may be an indirect consequence of its ability to suppress nonspecific binding of RNA polymerase II to DNA.

MeSH Terms
Base Sequence Cloning, Molecular DNA/metabolism Escherichia coli/enzymology,genetics,metabolism Glutathione Transferase/metabolism Humans Molecular Sequence Data RNA Polymerase II/metabolism Recombinant Proteins/metabolism Substrate Specificity Transcription Factors/isolation & purification,metabolism Transcription Factors, TFII Transcription, Genetic
Chemicals
Recombinant Proteins Transcription Factors Transcription Factors, TFII DNA Glutathione Transferase RNA Polymerase II transcription factor TFIIF
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Killeen M T
Department of Molecular and Medical Genetics, University of Toronto, Ontario, Canada.
Greenblatt J F
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46 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.1.30-37.1992
Published
1992-01-00
Pages
30-7
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC364066
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.12.1.30-37.1992 PMC Full Text View journal details
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