|Citation||Bielinsky, A.K., Blitzblau, H., Beall, E.L., Ezrokhi, M., Smith, H.S., Botchan, M.R., Gerbi, S.A. (2001). Origin recognition complex binding to a metazoan replication origin. Curr. Biol. 11(18): 1427--1431. (Export to RIS)|
|Publication Type||Research paper|
|PubMed Abstract||The initiation of DNA replication in eukaryotic cells at the onset of S phase requires the origin recognition complex (ORC) . This six-subunit complex, first isolated in Saccharomyces cerevisiae , is evolutionarily conserved . ORC participates in the formation of the prereplicative complex , which is necessary to establish replication competence. The ORC-DNA interaction is well established for autonomously replicating sequence (ARS) elements in yeast in which the ARS consensus sequence  (ACS) constitutes part of the ORC binding site [2, 5]. Little is known about the ORC-DNA interaction in metazoa. For the Drosophila chorion locus, it has been suggested that ORC binding is dispersed . We have analyzed the amplification origin (ori) II/9A of the fly, Sciara coprophila. We identified a distinct 80-base pair (bp) ORC binding site and mapped the replication start site located adjacent to it. The binding of ORC to this 80-bp core region is ATP dependent and is necessary to establish further interaction with an additional 65-bp of DNA. This is the first time that both the ORC binding site and the replication start site have been identified in a metazoan amplification origin. Thus, our findings extend the paradigm from yeast ARS1 to multicellular eukaryotes, implicating ORC as a determinant of the position of replication initiation.|
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|Language of Publication||English|
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