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Concerted Loading of Mcm2-7 Double Hexamers around DNA during DNA Replication Origin Licensing

Journal Article


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. © 2009 Elsevier Inc. All rights reserved.

Publication Date


  • 2009

Published In


Citation


  • Remus, D., Beuron, F., Tolun, G., Griffith, J. D., Morris, E. P., & Diffley, J. F. X. (2009). Concerted Loading of Mcm2-7 Double Hexamers around DNA during DNA Replication Origin Licensing. Cell, 139(4), 719-730. doi:10.1016/j.cell.2009.10.015

Scopus Eid


  • 2-s2.0-70350751416

Start Page


  • 719

End Page


  • 730

Volume


  • 139

Issue


  • 4

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. © 2009 Elsevier Inc. All rights reserved.

Publication Date


  • 2009

Published In


Citation


  • Remus, D., Beuron, F., Tolun, G., Griffith, J. D., Morris, E. P., & Diffley, J. F. X. (2009). Concerted Loading of Mcm2-7 Double Hexamers around DNA during DNA Replication Origin Licensing. Cell, 139(4), 719-730. doi:10.1016/j.cell.2009.10.015

Scopus Eid


  • 2-s2.0-70350751416

Start Page


  • 719

End Page


  • 730

Volume


  • 139

Issue


  • 4