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T-antigen-DNA polymerase alpha complex implicated in simian virus 40 DNA replication.

S T Smale·R Tjian
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Abstract

We have combined in vitro DNA replication reactions and immunological techniques to analyze biochemical interactions between simian virus (SV40) large T antigen and components of the cellular replication apparatus. First, in vitro SV40 DNA replication was characterized with specific origin mutants. Next, monoclonal antibodies were used to demonstrate that a specific domain of T antigen formed a complex with cellular DNA polymerase alpha. Several antibodies were identified that coprecipitated T antigen and DNA polymerase alpha, while others were found to selectively prevent this interaction and concomitantly inhibit DNA replication. DNA polymerase alpha also bound efficiently to a T-antigen affinity column, confirming the immunoprecipitation results and providing a useful method for purification of the complete protein complex. Taken together, these results suggest that the T-antigen-polymerase association may be a key step in the initiation of SV40 DNA replication.

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Antibodies, MonoclonalAntigen-Antibody ComplexAntigens, Polyomavirus TransformingAntigens, Viral, TumorDNA Polymerase IIDNA ReplicationHeLa CellsHumansMacromolecular SubstancesMutationOncogene Proteins, ViralProtein KinasesSimian virus 40Templates, GeneticJournal ArticleResearch Support, U.S. Gov't, Non-P.H.S.Research Support, U.S. Gov't, P.H.S.

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BibTeXRIS

S T Smale, R Tjian. 1986. T-antigen-DNA polymerase alpha complex implicated in simian virus 40 DNA replication.. https://doi.org/10.1128/mcb.6.11.4077-4087.1986

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