Interaction of DNA polymerase alpha-primase with cellular replication protein A and SV40 T antigen

EMBO J. 1992 Feb;11(2):769-76. doi: 10.1002/j.1460-2075.1992.tb05110.x.

Abstract

The purified human single-stranded DNA binding protein, replication protein A (RP-A), forms specific complexes with purified SV40 large T antigen and with purified DNA polymerase alpha-primase, as shown by ELISA and a modified immunoblotting technique. RP-A associated efficiently with the isolated primase, as well as with intact polymerase alpha-primase. The 70 kDa subunit of RP-A was sufficient for association with polymerase alpha-primase. Purified SV40 large T antigen bound to intact RP-A and to polymerase-primase, but not to any of the separated subunits of RP-A or to the isolated primase. These results suggest that the specific protein-protein interactions between RP-A, polymerase-primase and T antigen may play a role in the initiating of SV40 DNA replication.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal
  • Antigens, Polyomavirus Transforming / metabolism*
  • Cattle
  • DNA Primase
  • DNA-Binding Proteins / metabolism*
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli / metabolism*
  • Immunoblotting
  • Kinetics
  • Macromolecular Substances
  • Models, Structural
  • Molecular Sequence Data
  • Protein Binding
  • Protein Conformation
  • RNA Nucleotidyltransferases / isolation & purification
  • RNA Nucleotidyltransferases / metabolism*
  • Replication Protein A
  • Simian virus 40 / metabolism*
  • Thymus Gland / enzymology

Substances

  • Antibodies, Monoclonal
  • Antigens, Polyomavirus Transforming
  • DNA-Binding Proteins
  • Macromolecular Substances
  • Replication Protein A
  • DNA Primase
  • RNA Nucleotidyltransferases