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Sequence analyses revealed that spOrc1, 2, and 5 subunits are highly conserved compared with their counterparts from\n            <jats:italic>S. cerevisiae<\/jats:italic>\n            ,\n            <jats:italic>Xenopus<\/jats:italic>\n            ,\n            <jats:italic>Drosophila<\/jats:italic>\n            , and human. In contrast, both spOrc3 and spOrc6 subunits are poorly conserved. As reported by Chuang and Kelly [(1999)\n            <jats:italic>Proc. Natl. Acad. Sci. USA<\/jats:italic>\n            96, 2656\u20132661], the C-terminal region of spOrc4 is also conserved whereas the N terminus uniquely contains repeats of a sequence that binds strongly to AT-rich DNA regions. Consistent with this, extraction of\n            <jats:italic>S. pombe<\/jats:italic>\n            chromatin with 1 M NaCl, or after DNase I treatment, yielded the six-subunit ORC, whereas extraction with 0.3 M resulted in five-subunit ORC lacking spOrc4p. The spORC can be reconstituted\n            <jats:italic>in vitro<\/jats:italic>\n            with all six recombinant subunits expressed in the rabbit reticulocyte system. The association of spOrc4p with the other subunits required the removal of DNA from reaction mixture by DNase I. This suggests that a strong interaction between spOrc4p and DNA can prevent the isolation of the six-subunit ORC. The unique DNA-binding properties of the spORC may contribute to our understanding of the sequence-specific recognition required for the initiation of DNA replication in\n            <jats:italic>S. pombe<\/jats:italic>\n            .\n          <\/jats:p>","DOI":"10.1073\/pnas.96.22.12367","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T14:41:48Z","timestamp":1027694508000},"page":"12367-12372","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":52,"title":["Identification and reconstitution of the origin recognition complex from\n            <i>Schizosaccharomyces pombe<\/i>"],"prefix":"10.1073","volume":"96","author":[{"given":"Kyeong-Yeop","family":"Moon","sequence":"first","affiliation":[{"name":"Program of Molecular Biology, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, Box 97, New York, NY 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