Identification of multiple cellular factors required for SV40 replication in vitro

Fairman, M. P., Prelich, G., Stillman, B. (1987) Identification of multiple cellular factors required for SV40 replication in vitro. Philos Trans R Soc Lond B Biol Sci, 317 (1187). pp. 495-505. ISSN 0962-8436

URL: http://www.ncbi.nlm.nih.gov/pubmed/2894684
DOI: 10.1098/rstb.1987.0076

Abstract

The replication of simian virus 40 has been studied by using cell-free extracts derived from human 293 cells. Fractionation of this extract has led to the identification of three fractions that are required for efficient DNA synthesis. Initial fractionation of the crude extract by phosphocellulose chromatography has produced two fractions, I and II, neither of which is able to support replication separately, but when they are combined, efficient synthesis is restored. Both fractions are required, with SV40 T antigen, for the formation of a presynthesis complex at the SV40 origin. The major replication enzymes, DNA polymerase, DNA primase and the topoisomerases I and II all reside in fraction II. Fraction I has been subdivided into two subfractions (A and B) by DEAE-cellulose chromatography. Fraction A is essential for replication and is required for presynthesis complex formation. Fraction B stimulates DNA replication and is only required at the elongation stage. This multicomponent system has provided the foundation for identification of individual components that are required for DNA replication in vitro.

Item Type: Paper
Uncontrolled Keywords: Antigens Polyomavirus Transforming analysis Cell Line Cell-Free System DNA Primase DNA Replication DNA Topoisomerases Type I metabolism DNA Topoisomerases Type II metabolism DNA-Directed DNA Polymerase metabolism Humans Kinetics RNA Nucleotidyltransferases metabolism Simian virus 40 genetics immunology Virus Replication
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
biotechnology > chromatography
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date Deposited: 06 Mar 2012 16:43
Last Modified: 20 Jun 2017 13:25
Related URLs:
URI: http://repository.cshl.edu/id/eprint/24974

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