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

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Virshup, DM; Kelly, TJ

Published Date

  • May 1989

Published In

Volume / Issue

  • 86 / 10

Start / End Page

  • 3584 - 3588

PubMed ID

  • 2542940

Pubmed Central ID

  • 2542940

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.86.10.3584

Language

  • eng

Conference Location

  • United States