Purification of the simian virus 40 (SV40) T antigen DNA-binding domain and characterization of its interactions with the SV40 origin

Woo S. Joo, Xuelian Luo, Deborah Denis, Henry Y. Kim, Godfrey J. Rainey, Clyde Jones, K. R. Sreekumar, Peter A. Bullock

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

To better define protein-DNA interactions at a eukaryotic origin, the domain of simian virus 40 (SV40) large T antigen that specifically interacts with the SV40 origin has been purified and its binding to DNA has been characterized. Evidence is presented that the affinity of the purified T antigen DNA-binding domain for the SV40 origin is comparable to that of the full-length T antigen. Furthermore, stable binding of the T antigen DNA- binding domain to the SV40 origin requires pairs of pentanucleotide recognition sites separated by approximately one turn of a DNA double helix and positioned in a head-to-head orientation. Although two pairs of pentanucleotides are present in the SV40 origin, footprinting and band shift experiments indicate that binding is limited to dimer formation on a single pair of pentanucleotides. Finally, it is demonstrated that the T antigen DNA- binding domain interacts poorly with single-stranded DNA.

Original languageEnglish (US)
Pages (from-to)3972-3985
Number of pages14
JournalJournal of Virology
Volume71
Issue number5
StatePublished - 1997

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Simian virus 40
DNA-binding domains
Viral Tumor Antigens
antigens
DNA
single-stranded DNA
Single-Stranded DNA
T-DNA
Proteins
proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Joo, W. S., Luo, X., Denis, D., Kim, H. Y., Rainey, G. J., Jones, C., ... Bullock, P. A. (1997). Purification of the simian virus 40 (SV40) T antigen DNA-binding domain and characterization of its interactions with the SV40 origin. Journal of Virology, 71(5), 3972-3985.

Purification of the simian virus 40 (SV40) T antigen DNA-binding domain and characterization of its interactions with the SV40 origin. / Joo, Woo S.; Luo, Xuelian; Denis, Deborah; Kim, Henry Y.; Rainey, Godfrey J.; Jones, Clyde; Sreekumar, K. R.; Bullock, Peter A.

In: Journal of Virology, Vol. 71, No. 5, 1997, p. 3972-3985.

Research output: Contribution to journalArticle

Joo, WS, Luo, X, Denis, D, Kim, HY, Rainey, GJ, Jones, C, Sreekumar, KR & Bullock, PA 1997, 'Purification of the simian virus 40 (SV40) T antigen DNA-binding domain and characterization of its interactions with the SV40 origin', Journal of Virology, vol. 71, no. 5, pp. 3972-3985.
Joo, Woo S. ; Luo, Xuelian ; Denis, Deborah ; Kim, Henry Y. ; Rainey, Godfrey J. ; Jones, Clyde ; Sreekumar, K. R. ; Bullock, Peter A. / Purification of the simian virus 40 (SV40) T antigen DNA-binding domain and characterization of its interactions with the SV40 origin. In: Journal of Virology. 1997 ; Vol. 71, No. 5. pp. 3972-3985.
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