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

D. M. Virshup, M. G. Kauffman, Thomas Kelly

Research output: Contribution to journalArticle

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 (PP2A(c)). 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/PP2A(c) 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.

Original languageEnglish (US)
Pages (from-to)3891-3898
Number of pages8
JournalEMBO Journal
Volume8
Issue number12
StatePublished - 1989

Fingerprint

Protein Phosphatase 2
Replication Protein C
DNA Replication
Viral Tumor Antigens
Chemical activation
Phosphorylation
DNA
Proteins
Phosphoamino Acids
Polyomavirus Transforming Antigens
Viral DNA
G1 Phase
Viral Proteins
Catalytic Domain
Cell Cycle
Cells
In Vitro Techniques

Keywords

  • cell cycle regulation
  • phosphatase
  • T antigen

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Activation of SV40 DNA replication in vitro by cellular protein phosphatase 2A. / Virshup, D. M.; Kauffman, M. G.; Kelly, Thomas.

In: EMBO Journal, Vol. 8, No. 12, 1989, p. 3891-3898.

Research output: Contribution to journalArticle

Virshup, DM, Kauffman, MG & Kelly, T 1989, 'Activation of SV40 DNA replication in vitro by cellular protein phosphatase 2A', EMBO Journal, vol. 8, no. 12, pp. 3891-3898.
Virshup, D. M. ; Kauffman, M. G. ; Kelly, Thomas. / Activation of SV40 DNA replication in vitro by cellular protein phosphatase 2A. In: EMBO Journal. 1989 ; Vol. 8, No. 12. pp. 3891-3898.
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