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PMID: 1337880 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Purification and functional characterization of bovine RP-A in an in vitro SV40 DNA replication system.

Chromosoma ·Vol. 102 ·No. 1 Suppl ·1992-00-00 ·Pages S52-9

Nasheuer HP, von Winkler D, Schneider C, Dornreiter I, Gilbert I, Fanning E

Abstract

The single-stranded DNA binding protein RP-A is required in SV40 DNA in vitro replication. The RP-A purified from calf thymus contains 4 polypeptides with molecular weights 70kDa, 53kDa, 32kDa, and 14kDa. The p70 subunit and its proteolysed form p53 are recognized by the monoclonal antibody 70C (Kenny et al. (1990)) and bind to ssDNA. The p70 and p32 subunits of bovine RP-A are phosphorylated by CDC2-cyclin B kinase. Bovine RP-A supports the origin dependent unwinding of SV40 DNA by T antigen. Furthermore, bovine RP-A can efficiently substitute for human RP-A in SV40 DNA replication in vitro. A modified blotting technique revealed that RP-A interacts specifically and directly with the p48 subunit of DNA polymerase alpha-primase complex.

MeSH Terms
Ammonium Sulfate Animals Antigens, Polyomavirus Transforming/metabolism CDC2 Protein Kinase/physiology Cattle Cell-Free System DNA Helicases/physiology DNA Polymerase II/metabolism DNA Primase DNA Replication/physiology DNA, Single-Stranded/metabolism DNA-Binding Proteins/isolation & purification,physiology Humans Phosphorylation RNA Nucleotidyltransferases/metabolism Replication Protein A Simian virus 40
Chemicals
Antigens, Polyomavirus Transforming DNA, Single-Stranded DNA-Binding Proteins RPA1 protein, human Replication Protein A CDC2 Protein Kinase DNA Primase RNA Nucleotidyltransferases DNA Polymerase II DNA Helicases Ammonium Sulfate
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Nasheuer H P
Institute for Biochemistry, LMU München, Federal Republic of Germany.
von Winkler D
Schneider C
Dornreiter I
Gilbert I
Fanning E
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Article Info
Journal
Chromosoma
Abbr.
Chromosoma
ISSN
0009-5915
DOI
10.1007/BF02451786
Published
1992-00-00
Pages
S52-9
Language
English
Region
Austria
NLM ID
2985138R
Subset
IM
External Links
PubMed source DOI: 10.1007/BF02451786 View journal details
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