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 journalArticlepeer-review

33 Scopus citations

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
DOIs
StatePublished - May 1997

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

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