Purified JC virus T antigen derived from insect cells preferentially interacts with binding site II of the viral core origin under replication conditions

Virology
B BollagR J Frisque

Abstract

The human polyomavirus JC virus (JCV) establishes persistent, asymptomatic infections in most individuals, but in severely immunocompromised hosts it may cause the fatal demyelinating brain disease progressive multifocal leukoencephalopathy. In cell culture JCV multiplies inefficiently and exhibits a narrow host range. This restricted behavior occurs, in part, at the level of DNA replication, which is regulated by JCV's multifunctional large tumor protein (TAg). To prepare purified JCV TAg (JCT) for biochemical analyses, the recombinant baculovirus B-JCT was generated by cotransfection of insect cells with wild-type baculovirus and the vector pVL-JCT(Int-) containing the JCT-coding sequence downstream of the efficient polyhedrin promoter. JCT expressed in infected cells was immunoaffinity purified using the anti-JCT monoclonal antibody PAb 2000. Characterization of the viral oncoprotein indicated that it exists in solution as a mixture of monomeric and oligomeric species. With the addition of ATP, the population of monomers decreased and that of hexamers and double hexamers increased. A DNA mobility shift assay indicated that origin binding occurred primarily with the double-hexamer form. A comparison of the specific DNA-bindin...Continue Reading

Citations

Aug 31, 2000·Annual Review of Biochemistry·S S Patel, K M Picha
May 2, 2014·The Plant Journal : for Cell and Molecular Biology·C Douglas GrubbSteffen Abel
Oct 22, 2014·Biochemistry and Molecular Biology Education : a Bimonthly Publication of the International Union of Biochemistry and Molecular Biology·Christopher W Carroll, Lani C Keller
May 6, 2016·Journal of Virology·Brandy VerhalenMengxi Jiang
Oct 6, 1997·Journal of Virology·J NesperH P Nasheuer

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