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| Subject Categories: Genome Stability & Dynamics |
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| The EMBO Journal
(2006) 25, 5372–5382, doi:10.1038/sj.emboj.7601396 Published online 26 October 2006 |
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| The BAH domain facilitates the ability of human Orc1 protein to activate replication origins in vivo |
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| Kohji Noguchi1, 3, Alex Vassilev1, Soma Ghosh1, John L Yates2 and Melvin L DePamphilis1 |
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1 National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA 2 Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, NY, USA To whom correspondence should be addressed Melvin L DePamphilis, National Institute of Child Health and Human Development, National Institutes of Health, Building 6/3A15, 9000 Rockville Pike, Bethesda, MD 20892-2753, USA. Tel.: +1 301 402 8234; Fax: +1 301 480 9354; E-mail: depamphm@mail.nih.gov 3 Present address: Department of Bioactive Molecules, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan Received 28 June 2006; Accepted 21 September 2006; Published online 26 October 2006. |
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| Abstract |
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| Selection of initiation sites for DNA replication in eukaryotes is determined by the interaction between the origin recognition complex (ORC) and genomic DNA. In mammalian cells, this interaction appears to be regulated by Orc1, the only ORC subunit that contains a bromo-adjacent homology (BAH) domain. Since BAH domains mediate protein–protein interactions, the human Orc1 BAH domain was mutated, and the mutant proteins expressed in human cells to determine their affects on ORC function. The BAH domain was not required for nuclear localization of Orc1, association of Orc1 with other ORC subunits, or selective degradation of Orc1 during S-phase. It did, however, facilitate reassociation of Orc1 with chromosomes during the M to G1-phase transition, and it was required for binding Orc1 to the Epstein–Barr virus oriP and stimulating oriP-dependent plasmid DNA replication. Moreover, the BAH domain affected Orc1's ability to promote binding of Orc2 to chromatin as cells exit mitosis. Thus, the BAH domain in human Orc1 facilitates its ability to activate replication origins in vivo by promoting association of ORC with chromatin. |
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| Keywords: cell cycle, DNA replication, Epstein–Barr virus, origin recognition complex, oriP |
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