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PMID: 1318194 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cell cycle regulated phosphorylation of RPA-32 occurs within the replication initiation complex.

The EMBO journal ·Vol. 11 ·No. 6 ·1992-06-00 ·Pages 2177-87

Fotedar R, Roberts JM

Abstract

The transition from G1 to S phase of the cell cycle may be regulated by modification of proteins which are essential for initiating DNA replication. One of the first events during initiation is to unwind the origin DNA and this requires a single-stranded DNA binding protein. RPA, a highly conserved multi-subunit single-stranded DNA binding protein, was first identified as a cellular protein necessary for the initiation of SV40 DNA replication. The 32 kDa subunit of RPA has been shown to be phosphorylated at the start of S phase. Using SV40 replication as a model, we have reproduced in vitro the S phase-dependent phosphorylation of RPA-32 and show that it occurs specifically within the replication initiation complex. Phosphorylated RPA-32 is predominantly associated with DNA. Phosphorylation is not a pre-requisite for association with DNA, but occurs after RPA binds to single-stranded DNA formed at the origin during the initiation phase. The protein kinase(s) which phosphorylates RPA-32 is present at all stages of the cell cycle but RPA-32 does not bind to the SV40 origin or become phosphorylated in extracts from G1 cells. Therefore, the cell cycle-dependent phosphorylation of RPA-32 may be regulated by its binding to single-stranded origin DNA during replication initiation.

MeSH Terms
Animals CDC2 Protein Kinase/metabolism Cell Cycle/physiology Cell Line DNA Helicases/metabolism DNA Replication DNA, Single-Stranded/genetics DNA, Viral/genetics G1 Phase/physiology Macromolecular Substances Peptide Mapping Phosphates/metabolism Phosphoprotein Phosphatases/metabolism Phosphorus Radioisotopes Phosphorylation Protein Kinases/metabolism S Phase/physiology Simian virus 40/genetics
Chemicals
DNA, Single-Stranded DNA, Viral Macromolecular Substances Phosphates Phosphorus Radioisotopes Protein Kinases CDC2 Protein Kinase Phosphoprotein Phosphatases DNA Helicases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Fotedar R
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
Roberts J M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1992-06-00
Pages
2177-87
Language
English
Region
England
NLM ID
8208664
PMCID
PMC556685
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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