Purification of replication protein C, a cellular protein involved in the initial stages of simian virus 40 DNA replication in vitro.

Abstract:

:The replication of simian virus 40 (SV40) DNA is dependent upon a single viral protein [tumor (T) antigen] and multiple cellular proteins. To define the required cellular proteins, we have made use of a cell-free system that supports the replication of plasmid DNA molecules containing the SV40 origin of replication. We report here the purification from HeLa cell extracts of replication protein C (RP-C), a previously undescribed protein that is required to reconstitute efficient DNA replication in vitro. Highly purified preparations of RP-C contain two closely related polypeptides of 32 and 34 kDa. Preincubation of purified RP-C with T antigen and the DNA template largely eliminates the delay normally observed before the onset of rapid DNA synthesis. In addition, RP-C stimulates the unwinding of duplex DNA molecules containing the SV40 replication origin in a reaction that requires T antigen and a single-stranded DNA binding protein. These observations suggest that RP-C is involved in the initial steps of SV40 DNA replication in vitro.

authors

Virshup DM,Kelly TJ

doi

10.1073/pnas.86.10.3584

subject

Has Abstract

pub_date

1989-05-01 00:00:00

pages

3584-8

issue

10

eissn

0027-8424

issn

1091-6490

journal_volume

86

pub_type

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