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PMID: 9882311 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Interaction of the transcription factor TFIID with simian virus 40 (SV40) large T antigen interferes with replication of SV40 DNA in vitro.

Journal of virology ·Vol. 73 ·No. 2 ·1999-02-00 ·Pages 1099-107

Herbig U, Weisshart K, Taneja P, Fanning E

Abstract

Simian virus 40 (SV40) large tumor (T) antigen is the major regulatory protein that directs the course of viral infection, primarily by interacting with host cell proteins and modulating their functions. Initiation of viral DNA replication requires specific interactions of T antigen bound to the viral origin of DNA replication with cellular replication proteins. Transcription factors are thought to stimulate initiation of viral DNA replication, but the mechanism of stimulation is poorly understood. Since the transcription factor TATA-binding protein (TBP) binds to sequences within the origin of replication and interacts specifically with T antigen, we examined whether TBP complexes stimulate SV40 DNA replication in vitro. On the contrary, we found that depletion of TBP complexes from human cell extracts increased their ability to support viral DNA replication, and readdition of TBP complexes to the depleted extracts diminished their activity. We have mapped the sites of interaction between the proteins to residues 181 to 205 of T antigen and 184 to 220 of TBP. Titration of fusion proteins containing either of these peptides into undepleted cell extracts stimulated their replication activity, suggesting that they prevented the T antigen-TBP interaction that interfered with replication activity. TBP complexes also interfered with origin DNA unwinding by purified T antigen, and addition of either the T antigen or the TBP fusion peptide relieved the inhibition. These results suggest that TBP complexes associate with a T-antigen surface that is also required for origin DNA unwinding and viral DNA replication. We speculate that competition among cellular proteins for T antigen may play a role in regulating the course of viral infection.

MeSH Terms
Animals Antigens, Polyomavirus Transforming/genetics,metabolism Binding Sites DNA Replication DNA, Viral DNA-Binding Proteins/metabolism Humans Recombinant Fusion Proteins/genetics,metabolism Simian virus 40/genetics,physiology TATA-Box Binding Protein Transcription Factor TFIID Transcription Factors/metabolism Transcription Factors, TFII/metabolism Virus Replication
Chemicals
Antigens, Polyomavirus Transforming DNA, Viral DNA-Binding Proteins Recombinant Fusion Proteins TATA-Box Binding Protein Transcription Factor TFIID Transcription Factors Transcription Factors, TFII
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Herbig U
Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235, and Vanderbilt Cancer Center, Nashville, Tennessee 37232-6838, USA.
Weisshart K
Taneja P
Fanning E
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.73.2.1099-1107.1999
Published
1999-02-00
Pages
1099-107
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC103930
Subset
IM
Grants
NIGMS NIH HHS · R01 GM052948 · United States
NIGMS NIH HHS · GM 52948 · United States
External Links
PubMed source DOI: 10.1128/JVI.73.2.1099-1107.1999 PMC Full Text View journal details
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