Concerted Loading of Mcm2–7 Double Hexamers around DNA during DNA Replication Origin Licensing
The licensing of eukaryotic DNA replication origins, which ensures once-per-cell-cycle replication, involves the loading of six related minichromosome maintenance proteins (Mcm2–7) into prereplicative complexes (pre-RCs). Mcm2–7 forms the core of the replicative DNA helicase, which is inactive in th...
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| Veröffentlicht in: | Cell 2009-11, Vol.139 (4), p.719-730 |
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| creator | Remus, Dirk Beuron, Fabienne Tolun, Gökhan Griffith, Jack D. Morris, Edward P. Diffley, John F.X. |
| description | The licensing of eukaryotic DNA replication origins, which ensures once-per-cell-cycle replication, involves the loading of six related minichromosome maintenance proteins (Mcm2–7) into prereplicative complexes (pre-RCs). Mcm2–7 forms the core of the replicative DNA helicase, which is inactive in the pre-RC. The loading of Mcm2–7 onto DNA requires the origin recognition complex (ORC), Cdc6, and Cdt1, and depends on ATP. We have reconstituted Mcm2–7 loading with purified budding yeast proteins. Using biochemical approaches and electron microscopy, we show that single heptamers of Cdt1•Mcm2–7 are loaded cooperatively and result in association of stable, head-to-head Mcm2–7 double hexamers connected via their N-terminal rings. DNA runs through a central channel in the double hexamer, and, once loaded, Mcm2–7 can slide passively along double-stranded DNA. Our work has significant implications for understanding how eukaryotic DNA replication origins are chosen and licensed, how replisomes assemble during initiation, and how unwinding occurs during DNA replication. |
| doi_str_mv | 10.1016/j.cell.2009.10.015 |
| format | Article |
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Our work has significant implications for understanding how eukaryotic DNA replication origins are chosen and licensed, how replisomes assemble during initiation, and how unwinding occurs during DNA replication.</description><subject>Cell Cycle Proteins - chemistry</subject><subject>Cell Cycle Proteins - isolation & purification</subject><subject>Cell Cycle Proteins - metabolism</subject><subject>Chromosomal Proteins, Non-Histone - chemistry</subject><subject>Chromosomal Proteins, Non-Histone - isolation & purification</subject><subject>Chromosomal Proteins, Non-Histone - metabolism</subject><subject>DNA</subject><subject>DNA - metabolism</subject><subject>DNA Helicases - metabolism</subject><subject>DNA Replication</subject><subject>DNA-Binding Proteins - chemistry</subject><subject>DNA-Binding Proteins - isolation & purification</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Minichromosome Maintenance Complex Component 3</subject><subject>Minichromosome Maintenance Complex Component 4</subject><subject>Minichromosome Maintenance Complex Component 6</subject><subject>Minichromosome Maintenance Complex Component 7</subject><subject>Models, Molecular</subject><subject>Nuclear Proteins - chemistry</subject><subject>Nuclear Proteins - isolation & purification</subject><subject>Nuclear Proteins - metabolism</subject><subject>Origin Recognition Complex - metabolism</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - metabolism</subject><subject>Saccharomyces cerevisiae Proteins - chemistry</subject><subject>Saccharomyces cerevisiae Proteins - isolation & purification</subject><subject>Saccharomyces cerevisiae Proteins - metabolism</subject><issn>0092-8674</issn><issn>1097-4172</issn><issn>1097-4172</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkc2KFDEUhYMoTjv6Ai4kK11Ve5PKX4EIQ486QuuA6DpmUrfbNFWVnqRq0J3v4Bv6JKboxp_N4Crh3u-cm9xDyGMGSwZMPd8tPXbdkgM0pbAEJu-QBYNGV4JpfpcsSoNXRmlxQh7kvAMAI6W8T05YYxrFDF-Qz6s4eEwjtnQdXRuGLY0b-s73_Of3H5qex-mqQ3qBX12PKVOX4jS09Pz9GW2nNNPz9QPuu-DdGOJAL1PYhoGug8chF-AhubdxXcZHx_OUfHr96uPqolpfvnm7OltXXho-VlJy0SjPpNBSm8bIxqFRzsnWSacMOpBQO-5QMQQPDAXzSrUS0EDjwNen5OXBdz9d9diW6WNynd2n0Lv0zUYX7L-dIXyx23hjuQFhpCkGz44GKV5PmEfbhzwv2A0Yp2x1LZhgTOhCPr2V5IwzXav6P0AwWgIUkB9An2LOCTe_383AzlnbnZ11ds56rpWsi-jJ3z_-IzmGW4AXBwDL3m8CJpt9wBJ3GxL60bYx3Ob_CzzmusQ</recordid><startdate>20091113</startdate><enddate>20091113</enddate><creator>Remus, Dirk</creator><creator>Beuron, Fabienne</creator><creator>Tolun, Gökhan</creator><creator>Griffith, Jack D.</creator><creator>Morris, Edward P.</creator><creator>Diffley, John F.X.</creator><general>Elsevier Inc</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20091113</creationdate><title>Concerted Loading of Mcm2–7 Double Hexamers around DNA during DNA Replication Origin Licensing</title><author>Remus, Dirk ; Beuron, Fabienne ; Tolun, Gökhan ; Griffith, Jack D. ; Morris, Edward P. ; Diffley, John F.X.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c582t-552496c15475789859ae86aa5da5a68ea0503a2ae61e0c01e41c66d50e809a0c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Cell Cycle Proteins - chemistry</topic><topic>Cell Cycle Proteins - isolation & purification</topic><topic>Cell Cycle Proteins - metabolism</topic><topic>Chromosomal Proteins, Non-Histone - chemistry</topic><topic>Chromosomal Proteins, Non-Histone - isolation & purification</topic><topic>Chromosomal Proteins, Non-Histone - metabolism</topic><topic>DNA</topic><topic>DNA - metabolism</topic><topic>DNA Helicases - metabolism</topic><topic>DNA Replication</topic><topic>DNA-Binding Proteins - chemistry</topic><topic>DNA-Binding Proteins - isolation & purification</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Minichromosome Maintenance Complex Component 3</topic><topic>Minichromosome Maintenance Complex Component 4</topic><topic>Minichromosome Maintenance Complex Component 6</topic><topic>Minichromosome Maintenance Complex Component 7</topic><topic>Models, Molecular</topic><topic>Nuclear Proteins - chemistry</topic><topic>Nuclear Proteins - isolation & purification</topic><topic>Nuclear Proteins - metabolism</topic><topic>Origin Recognition Complex - metabolism</topic><topic>Saccharomyces cerevisiae</topic><topic>Saccharomyces cerevisiae - metabolism</topic><topic>Saccharomyces cerevisiae Proteins - chemistry</topic><topic>Saccharomyces cerevisiae Proteins - isolation & purification</topic><topic>Saccharomyces cerevisiae Proteins - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Remus, Dirk</creatorcontrib><creatorcontrib>Beuron, Fabienne</creatorcontrib><creatorcontrib>Tolun, Gökhan</creatorcontrib><creatorcontrib>Griffith, Jack D.</creatorcontrib><creatorcontrib>Morris, Edward P.</creatorcontrib><creatorcontrib>Diffley, John F.X.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Cell</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Remus, Dirk</au><au>Beuron, Fabienne</au><au>Tolun, Gökhan</au><au>Griffith, Jack D.</au><au>Morris, Edward P.</au><au>Diffley, John F.X.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Concerted Loading of Mcm2–7 Double Hexamers around DNA during DNA Replication Origin Licensing</atitle><jtitle>Cell</jtitle><addtitle>Cell</addtitle><date>2009-11-13</date><risdate>2009</risdate><volume>139</volume><issue>4</issue><spage>719</spage><epage>730</epage><pages>719-730</pages><issn>0092-8674</issn><issn>1097-4172</issn><eissn>1097-4172</eissn><abstract>The licensing of eukaryotic DNA replication origins, which ensures once-per-cell-cycle replication, involves the loading of six related minichromosome maintenance proteins (Mcm2–7) into prereplicative complexes (pre-RCs). Mcm2–7 forms the core of the replicative DNA helicase, which is inactive in the pre-RC. The loading of Mcm2–7 onto DNA requires the origin recognition complex (ORC), Cdc6, and Cdt1, and depends on ATP. We have reconstituted Mcm2–7 loading with purified budding yeast proteins. Using biochemical approaches and electron microscopy, we show that single heptamers of Cdt1•Mcm2–7 are loaded cooperatively and result in association of stable, head-to-head Mcm2–7 double hexamers connected via their N-terminal rings. DNA runs through a central channel in the double hexamer, and, once loaded, Mcm2–7 can slide passively along double-stranded DNA. Our work has significant implications for understanding how eukaryotic DNA replication origins are chosen and licensed, how replisomes assemble during initiation, and how unwinding occurs during DNA replication.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>19896182</pmid><doi>10.1016/j.cell.2009.10.015</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete; Cell Press Free Archives; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals |
| subjects | Cell Cycle Proteins - chemistry Cell Cycle Proteins - isolation & purification Cell Cycle Proteins - metabolism Chromosomal Proteins, Non-Histone - chemistry Chromosomal Proteins, Non-Histone - isolation & purification Chromosomal Proteins, Non-Histone - metabolism DNA DNA - metabolism DNA Helicases - metabolism DNA Replication DNA-Binding Proteins - chemistry DNA-Binding Proteins - isolation & purification DNA-Binding Proteins - metabolism Minichromosome Maintenance Complex Component 3 Minichromosome Maintenance Complex Component 4 Minichromosome Maintenance Complex Component 6 Minichromosome Maintenance Complex Component 7 Models, Molecular Nuclear Proteins - chemistry Nuclear Proteins - isolation & purification Nuclear Proteins - metabolism Origin Recognition Complex - metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae - metabolism Saccharomyces cerevisiae Proteins - chemistry Saccharomyces cerevisiae Proteins - isolation & purification Saccharomyces cerevisiae Proteins - metabolism |
| title | Concerted Loading of Mcm2–7 Double Hexamers around DNA during DNA Replication Origin Licensing |
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