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

Publication ,  Journal Article
Virshup, DM; Kelly, TJ
Published in: Proc Natl Acad Sci U S A
May 1989

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.

Duke Scholars

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

May 1989

Volume

86

Issue

10

Start / End Page

3584 / 3588

Location

United States

Related Subject Headings

  • Virus Replication
  • Viral Proteins
  • Simian virus 40
  • Molecular Weight
  • Kinetics
  • In Vitro Techniques
  • Hela Cells
  • HeLa Cells
  • DNA Replication
  • DNA Helicases
 

Citation

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Virshup, D. M., & Kelly, T. J. (1989). Purification of replication protein C, a cellular protein involved in the initial stages of simian virus 40 DNA replication in vitro. Proc Natl Acad Sci U S A, 86(10), 3584–3588. https://doi.org/10.1073/pnas.86.10.3584
Virshup, D. M., and T. J. Kelly. “Purification of replication protein C, a cellular protein involved in the initial stages of simian virus 40 DNA replication in vitro.Proc Natl Acad Sci U S A 86, no. 10 (May 1989): 3584–88. https://doi.org/10.1073/pnas.86.10.3584.
Virshup, D. M., and T. J. Kelly. “Purification of replication protein C, a cellular protein involved in the initial stages of simian virus 40 DNA replication in vitro.Proc Natl Acad Sci U S A, vol. 86, no. 10, May 1989, pp. 3584–88. Pubmed, doi:10.1073/pnas.86.10.3584.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

May 1989

Volume

86

Issue

10

Start / End Page

3584 / 3588

Location

United States

Related Subject Headings

  • Virus Replication
  • Viral Proteins
  • Simian virus 40
  • Molecular Weight
  • Kinetics
  • In Vitro Techniques
  • Hela Cells
  • HeLa Cells
  • DNA Replication
  • DNA Helicases