RECOGNITION OF THE ADENOVIRUS TYPE-2 ORIGIN OF DNA-REPLICATION BY THE VIRALLY ENCODED DNA-POLYMERASE AND PRETERMINAL PROTEINS

被引:63
作者
TEMPERLEY, SM [1 ]
HAY, RT [1 ]
机构
[1] UNIV ST ANDREWS,DEPT BIOCHEM & MICROBIOL,IRVINE BLDG,NORTH ST,ST ANDREWS KY16 9AL,FIFE,SCOTLAND
关键词
ADENOVIRUS; DNA REPLICATION; DNA PROTEIN INTERACTION; DNA POLYMERASE; PRETERMINAL PROTEIN;
D O I
10.1002/j.1460-2075.1992.tb05109.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
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页码:761 / 768
页数:8
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