Reconstitution of simian virus 40 DNA replication with purified proteins.

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, a...

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Main Authors: Weinberg, D H, Collins, K L, Simancek, P, Russo, A, Wold, M S, Virshup, D M, Kelly, T J
Formato: Artigo
Idioma:en
Publicado em: 1990
Assuntos:
Research Article
Acesso em linha:https://ncbi.nlm.nih.gov/pmc/articles/PMC55025/
https://ncbi.nlm.nih.gov/pubmed/2174160
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spelling pubmed-550252001-08-25 Reconstitution of simian virus 40 DNA replication with purified proteins. Weinberg, D H Collins, K L Simancek, P Russo, A Wold, M S Virshup, D M Kelly, T J Proc Natl Acad Sci U S A Research 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. 1990-11 /pmc/articles/PMC55025/ /pubmed/2174160 Text en
institution US National Library of Medicine
collection PubMed Central
language en
format Article
topic Research Article
spellingShingle Research Article
Weinberg, D H
Collins, K L
Simancek, P
Russo, A
Wold, M S
Virshup, D M
Kelly, T J
Reconstitution of simian virus 40 DNA replication with purified proteins.
description 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.
author Weinberg, D H
Collins, K L
Simancek, P
Russo, A
Wold, M S
Virshup, D M
Kelly, T J
author_facet Weinberg, D H
Collins, K L
Simancek, P
Russo, A
Wold, M S
Virshup, D M
Kelly, T J
author_sort Weinberg, D H
title Reconstitution of simian virus 40 DNA replication with purified proteins.
title_short Reconstitution of simian virus 40 DNA replication with purified proteins.
title_full Reconstitution of simian virus 40 DNA replication with purified proteins.
title_fullStr Reconstitution of simian virus 40 DNA replication with purified proteins.
title_full_unstemmed Reconstitution of simian virus 40 DNA replication with purified proteins.
title_sort reconstitution of simian virus 40 dna replication with purified proteins.
publishDate 1990
url https://ncbi.nlm.nih.gov/pmc/articles/PMC55025/
https://ncbi.nlm.nih.gov/pubmed/2174160
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