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Reconstitution of simian virus 40 DNA replication with purified proteins.

Publication ,  Journal Article
Weinberg, DH; Collins, KL; Simancek, P; Russo, A; Wold, MS; Virshup, DM; Kelly, TJ
Published in: Proc Natl Acad Sci U S A
November 1990
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Replication of plasmid DNA molecules containing the simian virus 40 (SV40) origin of DNA replication has been reconstituted with seven highly purified cellular proteins plus the SV40 large tumor (T) antigen. Initiation of DNA synthesis is absolutely dependent upon T antigen, replication protein A, and the DNA polymerase alpha-primase complex and is stimulated by the catalytic subunit of protein phosphatase 2A. Efficient elongation of nascent chains additionally requires proliferating cell nuclear antigen, replication factor C, DNA topoisomerase I, and DNA polymerase delta. Electron microscopic studies indicate that DNA replication begins at the viral origin and proceeds via intermediates containing two forks that move in opposite directions. These findings indicate that the reconstituted replication reaction has many of the characteristics expected of authentic viral DNA replication.

Duke Scholars

David Marc Virshup
Author David Marc Virshup Pediatrics
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Published In

Proc Natl Acad Sci U S A

DOI

10.1073/pnas.87.22.8692

ISSN

0027-8424

Publication Date

November 1990

Volume

87

Issue

22

Start / End Page

8692 / 8696

Location

United States

Related Subject Headings

  • Simian virus 40
  • RNA, Viral
  • Proteins
  • Microscopy, Electron
  • In Vitro Techniques
  • DNA Replication
  • Cell-Free System
  • Antigens, Polyomavirus Transforming
 

Citation

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Weinberg, D. H., Collins, K. L., Simancek, P., Russo, A., Wold, M. S., Virshup, D. M., & Kelly, T. J. (1990). Reconstitution of simian virus 40 DNA replication with purified proteins. Proc Natl Acad Sci U S A, 87(22), 8692–8696. https://doi.org/10.1073/pnas.87.22.8692
Weinberg, D. H., K. L. Collins, P. Simancek, A. Russo, M. S. Wold, D. M. Virshup, and T. J. Kelly. “Reconstitution of simian virus 40 DNA replication with purified proteins.” Proc Natl Acad Sci U S A 87, no. 22 (November 1990): 8692–96. https://doi.org/10.1073/pnas.87.22.8692.
Weinberg DH, Collins KL, Simancek P, Russo A, Wold MS, Virshup DM, et al. Reconstitution of simian virus 40 DNA replication with purified proteins. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8692–6.
Weinberg, D. H., et al. “Reconstitution of simian virus 40 DNA replication with purified proteins.” Proc Natl Acad Sci U S A, vol. 87, no. 22, Nov. 1990, pp. 8692–96. Pubmed, doi:10.1073/pnas.87.22.8692.
Weinberg DH, Collins KL, Simancek P, Russo A, Wold MS, Virshup DM, Kelly TJ. Reconstitution of simian virus 40 DNA replication with purified proteins. Proc Natl Acad Sci U S A. 1990 Nov;87(22):8692–8696.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

10.1073/pnas.87.22.8692

ISSN

0027-8424

Publication Date

November 1990

Volume

87

Issue

22

Start / End Page

8692 / 8696

Location

United States

Related Subject Headings

  • Simian virus 40
  • RNA, Viral
  • Proteins
  • Microscopy, Electron
  • In Vitro Techniques
  • DNA Replication
  • Cell-Free System
  • Antigens, Polyomavirus Transforming