Reconstitution of simian virus 40 DNA replication with purified proteins.

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Weinberg, DH; Collins, KL; Simancek, P; Russo, A; Wold, MS; Virshup, DM; Kelly, TJ

Published Date

  • November 1990

Published In

Volume / Issue

  • 87 / 22

Start / End Page

  • 8692 - 8696

PubMed ID

  • 2174160

Pubmed Central ID

  • 2174160

Electronic International Standard Serial Number (EISSN)

  • 1091-6490

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.87.22.8692

Language

  • eng