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PMID: 2555176 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Activation of SV40 DNA replication in vitro by cellular protein phosphatase 2A.

The EMBO journal ·Vol. 8 ·No. 12 ·1989-12-01 ·Pages 3891-8

Virshup DM, Kauffman MG, Kelly TJ

Abstract

We have made use of the cell-free SV40 DNA replication system to identify and characterize cellular proteins required for efficient DNA synthesis. One such protein, replication protein C (RP-C), was shown to be involved with SV40 large T antigen in the early stages of viral DNA replication in vitro. We demonstrate here that RP-C is identical to the catalytic subunit of cellular protein phosphatase 2A (PP2Ac). The purified protein dephosphorylates specific phosphoamino acid residues in T antigen, consistent with the hypothesis that SV40 DNA replication is regulated by modulating the phosphorylation state of the viral initiator protein. We also show that purified RP-C/PP2Ac preferentially stimulates SV40 DNA replication in extracts from early G1 phase cells. This finding suggests that the activity of a cellular factor that influences the net phosphorylation state of T antigen is cell cycle dependent.

MeSH Terms
Alkaline Phosphatase/metabolism Amino Acid Sequence Antigens, Viral, Tumor/metabolism Catalysis DNA Replication DNA, Viral/biosynthesis HeLa Cells Humans Interphase Molecular Sequence Data Phosphoprotein Phosphatases/physiology Phosphorylation Protein Phosphatase 2 Simian virus 40/genetics
Chemicals
Antigens, Viral, Tumor DNA, Viral Alkaline Phosphatase Phosphoprotein Phosphatases Protein Phosphatase 2
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Virshup D M
Department of Molecular Biology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
Kauffman M G
Kelly T J
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1989-12-01
Pages
3891-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC402079
Subset
IM
Grants
NCI NIH HHS · CA-16519 · United States
NCI NIH HHS · CA-40414 · United States
NIDDK NIH HHS · DK-01528 · United States
Databases
GENBANK
J03804, J03805
External Links
PubMed source PMC Full Text View journal details
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