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PMID: 10954562 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Phosphorylation of simian virus 40 T antigen on Thr 124 selectively promotes double-hexamer formation on subfragments of the viral core origin.

Journal of virology ·Vol. 74 ·No. 18 ·2000-09-00 ·Pages 8601-13

Barbaro BA, Sreekumar KR, Winters DR, Prack AE, Bullock PA

Abstract

Cell cycle-dependent phosphorylation of simian virus 40 (SV40) large tumor antigen (T-ag) on threonine 124 is essential for the initiation of viral DNA replication. A T-ag molecule containing a Thr-->Ala substitution at this position (T124A) was previously shown to bind to the SV40 core origin but to be defective in DNA unwinding and initiation of DNA replication. However, exactly what step in the initiation process is defective as a result of the T124A mutation has not been established. Therefore, to better understand the control of SV40 replication, we have reinvestigated the assembly of T124A molecules on the SV40 origin. Herein it is demonstrated that hexamer formation is unaffected by the phosphorylation state of Thr 124. In contrast, T124A molecules are defective in double-hexamer assembly on subfragments of the core origin containing single assembly units. We also report that T124A molecules are inhibitors of T-ag double hexamer formation. These and related studies indicate that phosphorylation of T-ag on Thr 124 is a necessary step for completing the assembly of functional double hexamers on the SV40 origin. The implications of these studies for the cell cycle control of SV40 DNA replication are discussed.

MeSH Terms
Amino Acid Substitution Animals Antigens, Polyomavirus Transforming/chemistry,metabolism Base Sequence Models, Biological Molecular Sequence Data Mutation Oligonucleotides Peptide Fragments Phosphorylation Replication Origin Simian virus 40/physiology Threonine/metabolism Viral Core Proteins/chemistry,metabolism Virus Replication
Chemicals
Antigens, Polyomavirus Transforming Oligonucleotides Peptide Fragments Viral Core Proteins Threonine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Barbaro B A
Department of Biochemistry, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
Sreekumar K R
Winters D R
Prack A E
Bullock P A
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.74.18.8601-8613.2000
Published
2000-09-00
Pages
8601-13
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC116373
Subset
IM
Grants
NIGMS NIH HHS · R01 GM055397 · United States
NIGMS NIH HHS · 9RO1GM55397 · United States
External Links
PubMed source DOI: 10.1128/jvi.74.18.8601-8613.2000 PMC Full Text View journal details
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