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 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.

authors

Weinberg DH,Collins KL,Simancek P,Russo A,Wold MS,Virshup DM,Kelly TJ

doi

10.1073/pnas.87.22.8692

subject

Has Abstract

pub_date

1990-11-01 00:00:00

pages

8692-6

issue

22

eissn

0027-8424

issn

1091-6490

journal_volume

87

pub_type

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