Purification of replication protein C, a cellular protein involved in the initial stages of simian virus 40 DNA replication in vitro

D. M. Virshup, T. J. Kelly

Research output: Contribution to journalArticlepeer-review

Abstract

The replication of simian virus 40 (SV40) DNA is dependent upon a single viral protein [tumor (T) antigen] and multiple cellular proteins. To define the required cellular proteins, we have made use of a cell-free system that supports the replication of plasmid DNA molecules containing the SV40 origin of replication. We report here the purification from HeLa cell extracts of replication protein C (RP-C), a previously undescribed protein that is required to reconstitute efficient DNA replication in vitro. Highly purified preparations of RP-C contain two closely related polypeptides of 32 and 34 kDa. Preincubation of purified RP-C with T antigen and the DNA template largely eliminates the delay normally observed before the onset of rapid DNA synthesis. In addition, RP-C stimulates the unwinding of duplex DNA molecules containing the SV40 replication origin in a reaction that requires T antigen and a single-stranded DNA binding protein. These observations suggest that RP-C is involved in the initial steps of SV40 DNA replication in vitro.

Original languageEnglish (US)
Pages (from-to)3584-3588
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume86
Issue number10
DOIs
StatePublished - 1989

ASJC Scopus subject areas

  • General

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