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PMID: 9566887 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Assembly of T-antigen double hexamers on the simian virus 40 core origin requires only a subset of the available binding sites.

Molecular and cellular biology ·Vol. 18 ·No. 5 ·1998-05-00 ·Pages 2677-87

Joo WS, Kim HY, Purviance JD, Sreekumar KR, Bullock PA

Abstract

Initiation of simian virus 40 (SV40) DNA replication is dependent upon the assembly of two T-antigen (T-ag) hexamers on the SV40 core origin. To further define the oligomerization mechanism, the pentanucleotide requirements for T-ag assembly were investigated. Here, we demonstrate that individual pentanucleotides support hexamer formation, while particular pairs of pentanucleotides suffice for the assembly of T-ag double hexamers. Related studies demonstrate that T-ag double hexamers formed on "active pairs" of pentanucleotides catalyze a set of previously described structural distortions within the core origin. For the four-pentanucleotide-containing wild-type SV40 core origin, footprinting experiments indicate that T-ag double hexamers prefer to bind to pentanucleotides 1 and 3. Collectively, these experiments demonstrate that only two of the four pentanucleotides in the core origin are necessary for T-ag assembly and the induction of structural changes in the core origin. Since all four pentanucleotides in the wild-type origin are necessary for extensive DNA unwinding, we concluded that the second pair of pentanucleotides is required at a step subsequent to the initial assembly process.

MeSH Terms
Antigens, Viral, Tumor/metabolism Base Sequence Binding Sites DNA Footprinting DNA Replication Models, Genetic Models, Molecular Molecular Sequence Data Nucleic Acid Conformation Protein Binding Protein Conformation Replication Origin Simian virus 40/genetics
Chemicals
Antigens, Viral, Tumor
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Joo W S
Department of Biochemistry, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
Kim H Y
Purviance J D
Sreekumar K R
Bullock P A
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.18.5.2677
Published
1998-05-00
Pages
2677-87
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC110647
Subset
IM
Grants
NIGMS NIH HHS · 9RO1GM55397 · United States
External Links
PubMed source DOI: 10.1128/MCB.18.5.2677 PMC Full Text View journal details
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