Identification of cellular proteins required for simian virus 40 DNA replication

M. S WOLD; D. H WEINBERG; D. M VIRSHUP; J. J LI; T. J KELLY

doi:10.1016/S0021-9258(19)81684-8

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Identification of cellular proteins required for simian virus 40 DNA replication

Journal article

Open access

Peer reviewed

Identification of cellular proteins required for simian virus 40 DNA replication

M. S WOLD, D. H WEINBERG, D. M VIRSHUP, J. J LI and T. J KELLY

The Journal of biological chemistry, Vol.264(5), pp.2801-2809

1989

DOI: 10.1016/S0021-9258(19)81684-8

PMID: 2536723

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Abstract

Study of the proteins involved in DNA replication of a model system such as SV40 is a first step in understanding eukaryotic chromosomal replication. Using a cell-free system that is capable of replicating plasmid DNA molecules containing the SV40 origin of replication, we conducted a series of systematic fractionation-reconstitution experiments for the purpose of identifying and characterizing the cellular proteins involved in SV40 DNA replication. In addition to the one viral-encoded replication protein, T antigen, we have identified and begun to characterize at least six cellular components from a HeLa cytoplasmic extract that are absolutely required for SV40 DNA replication in vitro. These include: (i) two partially purified fractions, CF IC and CF IIA, and (ii) four proteins that have been purified to near homogeneity, replication protein-A, proliferating cell nuclear antigen, DNA polymerase alpha-primase complex, and topoisomerase (I and II). Replication protein-A is a multi-subunit protein that has single-stranded DNA binding activity and is required for a T antigen-dependent, origin-dependent unwinding reaction which may be an important early step in initiation of replication. Fraction CF IC can stimulate this unwinding reaction, suggesting that it also may function during initiation. Proliferating cell nuclear antigen, DNA polymerase alpha-primase, and CF IIA all appear to be involved in elongation of nascent chains.

Molecular Genetics

Fundamental and applied biological sciences. Psychology

Biological and medical sciences

Replication

Molecular and cellular biology

Details

Title: Subtitle: Identification of cellular proteins required for simian virus 40 DNA replication
Creators: M. S WOLD - Johns Hopkins univ. school medicine, dep. molecular biology genetics, Baltimore MD 21205, United States
D. H WEINBERG - Johns Hopkins univ. school medicine, dep. molecular biology genetics, Baltimore MD 21205, United States
D. M VIRSHUP - Johns Hopkins univ. school medicine, dep. molecular biology genetics, Baltimore MD 21205, United States
J. 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: The Journal of biological chemistry, Vol.264(5), pp.2801-2809
DOI: 10.1016/S0021-9258(19)81684-8
PMID: 2536723
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: American Society for Biochemistry and Molecular Biology; Bethesda, MD
Language: English
Date published: 1989
Academic Unit: Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984025283702771

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