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Recognition of the adenovirus type 2 origin of DNA replication by the virally encoded DNA polymerase and preterminal proteins.

1992; Springer Nature; Volume: 11; Issue: 2 Linguagem: Inglês

10.1002/j.1460-2075.1992.tb05109.x

1460-2075

S.M. Temperley, Ronald T. Hay,

CRISPR and Genetic Engineering

Research Article1 February 1992free access Recognition of the adenovirus type 2 origin of DNA replication by the virally encoded DNA polymerase and preterminal proteins. S.M. Temperley S.M. Temperley Department of Biochemistry and Microbiology, University of St Andrews, Fife, UK. Search for more papers by this author R.T. Hay R.T. Hay Department of Biochemistry and Microbiology, University of St Andrews, Fife, UK. Search for more papers by this author S.M. Temperley S.M. Temperley Department of Biochemistry and Microbiology, University of St Andrews, Fife, UK. Search for more papers by this author R.T. Hay R.T. Hay Department of Biochemistry and Microbiology, University of St Andrews, Fife, UK. Search for more papers by this author Author Information S.M. Temperley1 and R.T. Hay1 1Department of Biochemistry and Microbiology, University of St Andrews, Fife, UK. The EMBO Journal (1992)11:761-768https://doi.org/10.1002/j.1460-2075.1992.tb05109.x PDFDownload PDF of article text and main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Initiation of adenovirus DNA synthesis is preceded by the assembly of a nucleoprotein complex at the origin of DNA replication containing three viral proteins, preterminal protein, DNA polymerase and DNA binding protein, and two cellular proteins, nuclear factors I and III. While sequence specific interactions of the cellular proteins with their cognate sites in the origin of DNA replication are well characterized, the question of how the viral replication proteins recognize the origin has remained unanswered. Preterminal protein and DNA polymerase were therefore purified to homogeneity from recombinant baculovirus infected insect cells. Gel filtration demonstrated that while DNA polymerase existed in monomeric and dimeric forms, preterminal protein was predominantly monomeric and when combined the proteins formed a stable heterodimer. In a gel electrophoresis DNA binding assay each of the protein species recognized DNA within the origin of DNA replication with unique specificity. Competition analysis and DNase I protection experiments revealed that although each protein could recognize the origin, the heterodimer did so with enhanced specificity, protecting bases 8–17 from cleavage with the nuclease. Thus the highly conserved ‘core’ of the origin of DNA replication, present in all human adenoviruses, is recognized by the preterminal protein--DNA polymerase heterodimer. Previous ArticleNext Article Volume 11Issue 21 February 1992In this issue RelatedDetailsLoading ...