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PMID: 1698613 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

SV40 T antigen binds directly to the large subunit of purified DNA polymerase alpha.

The EMBO journal ·Vol. 9 ·No. 10 ·1990-10-00 ·Pages 3329-36

Dornreiter I, Höss A, Arthur AK, Fanning E

Abstract

Purified SV40 large T antigen and purified DNA polymerase alpha-primase form a complex detectable by ELISA and by a modified immunoblotting technique. The interaction is specific for the large catalytic subunit of polymerase alpha. The amino terminal 83 amino acids of T antigen are both necessary and sufficient for binding to the polymerase. However, antibody epitopes located in the carboxy terminal ATPase domain of T antigen are masked in the polymerase-T antigen complex, and complex formation is inhibited by an antibody directed against the carboxy terminus of T antigen, suggesting that this region of T antigen, though not required for binding, is in close proximity to the bound polymerase. The affinity of human DNA polymerase alpha for T antigen is approximately 10-fold greater than that of polymerase alpha from calf thymus, consistent with the interpretation that polymerase alpha is at least in part responsible for the primate-specific replication of SV40 DNA in vivo and in vitro. The results suggest that specific protein-protein interaction between DNA polymerase alpha and T antigen plays an important role in viral DNA replication.

MeSH Terms
Animals Antigens, Polyomavirus Transforming Binding Sites Cattle Chromatography, Affinity Chromatography, Ion Exchange DNA Polymerase II/immunology,isolation & purification,metabolism DNA Primase Enzyme-Linked Immunosorbent Assay Epitopes/analysis Immunoblotting Kinetics Macromolecular Substances RNA Nucleotidyltransferases/immunology,isolation & purification,metabolism Recombinant Proteins/metabolism Simian virus 40/immunology,metabolism Thymus Gland/enzymology
Chemicals
Antigens, Polyomavirus Transforming Epitopes Macromolecular Substances Recombinant Proteins DNA Primase RNA Nucleotidyltransferases DNA Polymerase II
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Dornreiter I
Institute for Biochemistry, Munich, FRG.
Höss A
Arthur A K
Fanning E
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1990-10-00
Pages
3329-36
Language
English
Region
England
NLM ID
8208664
PMCID
PMC552070
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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