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

被引：34

作者：

Joo, WS ^{[1
]}

Luo, XL ^{[1
]}

Denis, D ^{[1
]}

Kim, HY ^{[1
]}

Rainey, GJ ^{[1
]}

Jones, C ^{[1
]}

Sreekumar, KR ^{[1
]}

Bullock, PA ^{[1
]}

机构：

[1] TUFTS UNIV,SCH MED,DEPT BIOCHEM A703,BOSTON,MA 02111

来源：

JOURNAL OF VIROLOGY | 1997年 / 71卷 / 05期

关键词：

D O I：

10.1128/JVI.71.5.3972-3985.1997

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

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.

引用

页码：3972 / 3985

页数：14

共 92 条

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