Studies on the mechanism of DNA linking by Epstein-Barr virus nuclear antigen 1.
Epstein-Barr virus nuclear antigen 1 (EBNA1) can both bind to and link DNA. Dimers of EBNA1 bind specific sites, two clusters of which, the FR and DS, comprise the necessary cis-acting elements of the Epstein-Barr viral origin of plasmid replication. EBNA1-dimers can link FR and DS, looping out the intervening DNA. EBNA1 can also intermolecularly link DNAs to which it binds. Residues of EBNA1 that can mediate linking have been mapped to at least three, non-overlapping domains. These domains, when fused to the dimerization and DNA-binding domain of GAL4, can self-associate and thereby link DNAs bound site specifically by GAL4. Two disparate mechanisms could underlie self-association of linking domains: 1) linking domains could associate with other linking domains directly, or 2) linking domains could associate indirectly by binding to a common nucleic acid intermediate. We have found that EBNA1 can link DNA by each of these mechanisms, however, the linking domains associate directly with a greater apparent affinity than through a nonspecific nucleic acid intermediate.
Mackey
D
D
McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA. mackey@oncology.wisc.edu
Sugden
B
B
D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D019281:Dimerization; D019309:Epstein-Barr Virus Nuclear Antigens / Q000378:metabolism*; D011089:Polydeoxyribonucleotides / Q000378:metabolism; D011485:Protein Binding; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000378:metabolism
D004268:DNA-Binding Proteins; D019309:Epstein-Barr Virus Nuclear Antigens; C052130:GAL4 protein, S cerevisiae; D011089:Polydeoxyribonucleotides; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C031156:poly d(I-C); D004247:DNA; D003850:Deoxyribonuclease I; D012259:Ribonuclease, Pancreatic