Replication of SV40 in vitro using proteins derived from a human cell extract

Fairman, M. P., Prelich, G., Tsurimoto, T., Stillman, B. (1989) Replication of SV40 in vitro using proteins derived from a human cell extract. J Cell Sci Suppl, 12. pp. 161-9. ISSN 0269-3518



In the presence of large T antigen and plasmids containing a functional origin of replication, extracts from a human cell line will support multiple rounds of simian virus 40 (SV40) replication in vitro. Fractionation of this extract has led to the identification of several factors, some of which have been purified to homogeneity. The characterisation of these proteins has led to the separation of SV40 replication in vitro into multiple stages. Two proteins, the cell cycle-regulated proliferating cell nuclear antigen and replication factor-C, have been shown to be essential for coordinating leading and lagging strand synthesis in this system. Another protein, replication factor-A, is a multi-subunit protein of 70, 34 and 11K (K = 10(3) Mr) polypeptides which, because of its high affinity for DNA, is thought to function as a eukaryotic single-stranded DNA binding protein. Interactions between other cellular factors are also described that effect the initiation of DNA replication, but are not required in a more purified system. In addition a model for a hypothetical replication fork is described, which suggests a role for both alpha- and delta-polymerases in this system, and may be applicable to higher eukaryotes.

Item Type: Paper
Uncontrolled Keywords: Antigens Viral DNA Polymerase II metabolism DNA Primase DNA Replication DNA-Binding Proteins analysis Electrophoresis Polyacrylamide Gel Humans Nuclear Proteins analysis Proliferating Cell Nuclear Antigen RNA Nucleotidyltransferases metabolism Simian virus 40 physiology Virus Replication
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
organism description > virus
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date: 1989
Date Deposited: 06 Mar 2012 16:48
Last Modified: 20 Jun 2017 20:28

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