Journal article
Reconstitution of simian virus 40 DNA replication with purified proteins
Proceedings of the National Academy of Sciences - PNAS, Vol.87(22), pp.8692-8696
11/1990
DOI: 10.1073/pnas.87.22.8692
PMCID: PMC55025
PMID: 2174160
Abstract
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.
Details
- Title: Subtitle
- Reconstitution of simian virus 40 DNA replication with purified proteins
- Creators
- D H Weinberg - Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205Kathleen L Collins - Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205P Simancek - Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205Alicia Russo - Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205Marc S Wold - Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205David M Virshup - Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205T J Kelly - Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205
- Resource Type
- Journal article
- Publication Details
- Proceedings of the National Academy of Sciences - PNAS, Vol.87(22), pp.8692-8696
- DOI
- 10.1073/pnas.87.22.8692
- PMID
- 2174160
- PMCID
- PMC55025
- NLM abbreviation
- Proc Natl Acad Sci U S A
- ISSN
- 0027-8424
- eISSN
- 1091-6490
- Publisher
- National Academy of Sciences
- Language
- English
- Date published
- 11/1990
- Academic Unit
- Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984024509102771
Metrics
26 Record Views