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PMID: 1851853 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Mapping the transcriptional transactivation function of simian virus 40 large T antigen.

Journal of virology ·Vol. 65 ·No. 6 ·1991-06-00 ·Pages 2778-90

Zhu JY, Rice PW, Chamberlain M, Cole CN

Abstract

T antigen is able to transactivate gene expression from the simian virus 40 (SV40) late promoter and from several other viral and cellular promoters. Neither the mechanisms of transactivation by T antigen nor the regions of T antigen required for this activity have been determined. To address the latter point, we have measured the ability of a set of SV40 large T antigen mutants to stimulate gene expression in CV-1 monkey kidney cells from the SV40 late promoter and Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter. Transactivation, although reduced, was retained by an N-terminal 138-amino-acid fragment of T antigen. Mutants with alterations at various locations within the N-terminal 85 amino acids transactivated the RSV LTR promoter less well than did wild-type T antigen. Most of these were also partially defective in their ability to transactivate the SV40 late promoter. Two mutants with lesions in the DNA-binding domain that were unable to bind to SV40 DNA were completely defective for transactivation of both promoter, while a third mutant with a lesion in the DNA-binding domain which retained origin-binding activity transactivated both promoters as well as did wild-type T antigen. Only a low level of transactivation was seen with mutant T antigens which had lesions in or near the zinc finger region (amino acids 300 to 350). Mutations which caused defects in ATPase activity, host range/helper function, binding to p53, binding to the retinoblastoma susceptibility protein, or nuclear localization had little or no effect on transactivation. These results suggest that N-terminal portion of T antigen possesses an activation activity. The data are consistent with the idea that the overall conformation of T antigen is important for transactivation and that mutations in other regions that reduce or eliminate transactivation do so by altering the conformation or orientation of the N-terminal region so that its ability to interact with various targets is diminished or abolished.

MeSH Terms
Adenosine Triphosphatases/genetics Antigens, Viral, Tumor/genetics Avian Sarcoma Viruses/genetics Base Sequence DNA, Viral/immunology Gene Expression Regulation, Viral Mutagenesis Peptide Mapping Promoter Regions, Genetic Protein Conformation Repetitive Sequences, Nucleic Acid Simian virus 40/genetics,immunology
Chemicals
Antigens, Viral, Tumor DNA, Viral Adenosine Triphosphatases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Zhu J Y
Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03756.
Rice P W
Chamberlain M
Cole C N
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.65.6.2778-2790.1991
Published
1991-06-00
Pages
2778-90
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC240892
Subset
IM
Grants
NCI NIH HHS · CA-08835 · United States
NCI NIH HHS · CA-23018 · United States
NCI NIH HHS · CA-39253 · United States
External Links
PubMed source DOI: 10.1128/JVI.65.6.2778-2790.1991 PMC Full Text View journal details
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