title: T-antigen-DNA polymerase alpha complex implicated in simian virus 40 DNA replication.
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.
mesh_terms: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000952:Antigens, Polyomavirus Transforming; D000957:Antigens, Viral, Tumor / Q000276:immunology / Q000378:metabolism*; D004257:DNA Polymerase II / Q000378:metabolism*; D004261:DNA Replication; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D009154:Mutation; D009856:Oncogene Proteins, Viral / Q000276:immunology / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D013539:Simian virus 40 / Q000235:genetics*; D013698:Templates, Genetic
chemical_list: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000952:Antigens, Polyomavirus Transforming; D000957:Antigens, Viral, Tumor; D046911:Macromolecular Substances; D009856:Oncogene Proteins, Viral; D011494:Protein Kinases; D004257:DNA Polymerase II