Binding Studies of SV40 T-antigen to SV40 Binding Site II

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1985 Sep 25

PMID 2997717

Citations 5

Authors

P Gottlieb

M S Nasoff

E F FISHER

A M Walsh

M H Caruthers

Affiliations

Soon will be listed here.

Abstract

SV40 T-Antigen binding site II was synthesized, cloned and analyzed for its ability to bind purified SV40 T-antigen. We report the binding constant of T-antigen for isolated site II. Using a filter binding assay the calculated binding constant was 6-8 fold less efficient than site I previously reported. Binding constants were calculated using two methods. The first was a direct calculation using a protein titration curve (KD). The second was by the ratio of measured association and dissociation rates. Both methods gave similar constants. Protection studies with SV40 T-antigen on the T-antigen binding sites in the wild-type array demonstrated that the binding constants of site I and site II are similar to those calculated for the individual sites. These results demonstrate that SV40 T-antigen does not bind cooperatively to sites one and two as earlier believed and are in agreement with recent observations emanating from several laboratories.

Citing Articles

Sequence requirements for the assembly of simian virus 40 T antigen and the T-antigen origin binding domain on the viral core origin of replication.

Kim H, Barbaro B, Joo W, Prack A, Sreekumar K, Bullock P J Virol. 1999; 73(9):7543-55.

PMID: 10438844 PMC: 104281. DOI: 10.1128/JVI.73.9.7543-7555.1999.

Simian virus 40 (SV40) T antigen binds specifically to double-stranded DNA but not to single-stranded DNA or DNA/RNA hybrids containing the SV40 regulatory sequences.

Auborn K, Markowitz R, Wang E, Yu Y, Prives C J Virol. 1988; 62(6):2204-8.

PMID: 3367427 PMC: 253332. DOI: 10.1128/JVI.62.6.2204-2208.1988.

Identification of simian virus 40 T-antigen residues important for specific and nonspecific binding to DNA and for helicase activity.

Simmons D, Young W J Virol. 1990; 64(10):4858-65.

PMID: 2168972 PMC: 247975. DOI: 10.1128/JVI.64.10.4858-4865.1990.

Four major sequence elements of simian virus 40 large T antigen coordinate its specific and nonspecific DNA binding.

Simmons D, Loeber G, Tegtmeyer P J Virol. 1990; 64(5):1973-83.

PMID: 2157865 PMC: 249351. DOI: 10.1128/JVI.64.5.1973-1983.1990.

The DNA-binding properties of polyomavirus large T antigen are altered by ATP and other nucleotides.

Lorimer H, Wang E, Prives C J Virol. 1991; 65(2):687-99.

PMID: 1846192 PMC: 239808. DOI: 10.1128/JVI.65.2.687-699.1991.

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