Reconstitution of simian virus 40 DNA replication with purified proteins

David H. Weinberg; Kathleen L. Collins; Pamela Simancek; Alicia Russo; Marc S. Wold; David M. Virshup; Thomas J. Kelly

Reconstitution of simian virus 40 DNA replication with purified proteins

David H. Weinberg, Kathleen L. Collins, Pamela Simancek, Alicia Russo, Marc S. Wold, David M. Virshup, Thomas J. Kelly

Research output: Contribution to journal › Article › peer-review

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 α-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 δ. 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.

Original language	English (US)
Pages (from-to)	8692-8696
Number of pages	5
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	87
Issue number	22
State	Published - Nov 1990
Externally published	Yes

Keywords

DNA polymerase δ
Eukaryotic DNA replication
Replication factor C

ASJC Scopus subject areas

General

Cite this

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title = "Reconstitution of simian virus 40 DNA replication with purified proteins",

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 α-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 δ. 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.",

keywords = "DNA polymerase δ, Eukaryotic DNA replication, Replication factor C",

author = "Weinberg, {David H.} and Collins, {Kathleen L.} and Pamela Simancek and Alicia Russo and Wold, {Marc S.} and Virshup, {David M.} and Kelly, {Thomas J.}",

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T1 - Reconstitution of simian virus 40 DNA replication with purified proteins

AU - Weinberg, David H.

AU - Collins, Kathleen L.

AU - Simancek, Pamela

AU - Russo, Alicia

AU - Wold, Marc S.

AU - Virshup, David M.

AU - Kelly, Thomas J.

PY - 1990/11

Y1 - 1990/11

N2 - 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 α-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 δ. 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.

AB - 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 α-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 δ. 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.

KW - DNA polymerase δ

KW - Eukaryotic DNA replication

KW - Replication factor C

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Reconstitution of simian virus 40 DNA replication with purified proteins

Abstract

Keywords

ASJC Scopus subject areas

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