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Interaction of simian virus 40 large T-antigen with cellular DNA polymerase α: studies with various T-antigen mutants of SV40

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Summary

Simian virus 40 (SV40) large T antigen binds to host cell DNA polymerase α and the T-antigen-DNA polymerase α complex is implicated in the initiation of viral DNA replication. We have examined various SV40 T-antigen mutants to test the correspondence between viral DNA replication and T-antigen-DNA polymerase α complex formation. The various SV40 T-antigen mutants were used to either infect or transfect African green monkey kidney cell line CV-1, and at different time intervals we measured the production of T-antigen and host cell DNA polymerase α by radioimmunoassay, complex formation by a sandwich radioimmunoassay and the amount of viral DNA synthesis by dot-blot hybridization analysis. There was a good correlation between complex formation and viral DNA synthesis in lytic mutants of SV40. Poor complex formation and correspondingly lower DNA synthesis were observed in the non-viable mutants of SV40, even though significant amounts of T-antigen and DNA polymerase α were present. Our results substantiate the earlier findings of T-antigen-DNA polymerase α complex formation and establish the need for formation of this complex in promoting viral DNA synthesis.

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  1. Department of BiochemistryDepartment of BiochemistryDepartment of BiochemistryUniversity of Nebraska Medical Center, Omaha, Nebraska, USA

    J. M. Pistillo & J. K. Vishwanatha

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  1. J. M. Pistillo
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  2. J. K. Vishwanatha
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Pistillo, J.M., Vishwanatha, J.K. Interaction of simian virus 40 large T-antigen with cellular DNA polymerase α: studies with various T-antigen mutants of SV40. Archives of Virology 118, 113–125 (1991). https://doi.org/10.1007/BF01311307

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  • DOI: https://doi.org/10.1007/BF01311307

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