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Cdc6-induced conformational changes in ORC bound to origin DNA revealed by cryo-electron microscopy.

The eukaryotic origin recognition complex (ORC) interacts with and remodels origins of DNA replication prior to initiation in S phase. Here, we report a single-particle cryo-EM-derived structure of the supramolecular assembly comprising Saccharomyces cerevisiae ORC, the replication initiation factor Cdc6, and double-stranded ARS1 origin DNA in the presence of ATPγS. The six subunits of ORC are arranged as Orc1:Orc4:Orc5:Orc2:Orc3, with Orc6 binding to Orc2. Cdc6 binding changes the conformation of ORC, in particular reorienting the Orc1 N-terminal BAH domain. Segmentation of the 3D map of ORC-Cdc6 on DNA and docking with the crystal structure of the homologous archaeal Orc1/Cdc6 protein suggest an origin DNA binding model in which the DNA tracks along the interior surface of the crescent-like ORC. Thus, ORC bends and wraps the DNA. This model is consistent with the observation that binding of a single Cdc6 extends the ORC footprint on origin DNA from both ends.

Pubmed ID: 22405012 RIS Download

Mesh terms: Cell Cycle Proteins | Cryoelectron Microscopy | DNA-Binding Proteins | Models, Molecular | Origin Recognition Complex | Protein Conformation | Protein Subunits | Saccharomyces cerevisiae Proteins | Transcription Factors

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Associated grants

  • Agency: NIGMS NIH HHS, Id: R01 GM045436-20
  • Agency: Medical Research Council, Id: S10 RR025415
  • Agency: NCRR NIH HHS, Id: GM74985
  • Agency: NIGMS NIH HHS, Id: R01 GM074985-03
  • Agency: NIGMS NIH HHS, Id: MRC_MC_U120085811
  • Agency: Medical Research Council, Id: S10 RR025415-01
  • Agency: NCRR NIH HHS, Id: R01 GM074985-04S1
  • Agency: NIGMS NIH HHS, Id: R01 GM074985
  • Agency: NIGMS NIH HHS, Id: GM45436
  • Agency: NIGMS NIH HHS, Id: R01 GM045436
  • Agency: NIGMS NIH HHS, Id:

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