Initiation of simian virus 40 DNA replication in vitro: large-tumor-antigen- and origin-dependent unwinding of the template

Marc S Wold; Joachim J LI; T. J KELLY

doi:10.1073/pnas.84.11.3643

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Initiation of simian virus 40 DNA replication in vitro: large-tumor-antigen- and origin-dependent unwinding of the template

Journal article

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Initiation of simian virus 40 DNA replication in vitro: large-tumor-antigen- and origin-dependent unwinding of the template

Marc S Wold, Joachim J LI and T. J KELLY

Proceedings of the National Academy of Sciences - PNAS, Vol.84(11), pp.3643-3647

1987

DOI: 10.1073/pnas.84.11.3643

PMID: 3035543

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Abstract

Analysis of the kinetics of simian virus 40 (SV40) DNA replication in vitro demonstrated the existence of a slow presynthesis reaction that occurs prior to onset of extensive chain elongation and is dependent on a subset of the cellular proteins required for the complete replication reaction. When the presynthesis reaction is carried out in the presence of topoisomerase I, it is possible to detect extensive unwinding of the template DNA. This unwinding reaction is specific for templates that contain the wild-type SV40 origin of DNA replication and requires SV40 large tumor antigen (T antigen), ATP, and a protein fraction derived from HeLa cells. The required cellular protein may be a eukaryotic single-stranded-DNA-binding protein (SSB), since unwinding of the template is also observed when Escherichia coli SSB is substituted for the HeLa protein fraction. These observations suggest that during the initial stages of SV40 DNA replication, T antigen binds specifically to the viral origin and locally unwinds the DNA. This origin-dependent unwinding reaction is presumably a prerequisite for subsequent priming and elongation steps.

Molecular Genetics

Fundamental and applied biological sciences. Psychology

Biological and medical sciences

Replication

Molecular and cellular biology

Details

Title: Subtitle: Initiation of simian virus 40 DNA replication in vitro: large-tumor-antigen- and origin-dependent unwinding of the template
Creators: Marc S Wold - Johns Hopkins univ. school medicine, dep. molecular biology genetics, Baltimore MD 21205, United States
Joachim J LI - Johns Hopkins univ. school medicine, dep. molecular biology genetics, Baltimore MD 21205, United States
T. J KELLY - Johns Hopkins univ. school medicine, dep. molecular biology genetics, Baltimore MD 21205, United States
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.84(11), pp.3643-3647
DOI: 10.1073/pnas.84.11.3643
PMID: 3035543
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences; Washington, DC
Language: English
Date published: 1987
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984025396002771

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