Fig. 10: The hORC2-WHD in the hOCCM is reorganised upon DNA and hCDC6 binding.
a Structure of hORC2-WHD. The hORC2 (mauve) is stabilised in the open claw position by the presence of hCDC6 AAA+ ___domain (light grey), as compared with the closed claw conformations of hORC2-WHD in hORC2-5 (PDB: 7JPQ) and in DmOrc2 (PDB: 4XGC) which is not compatible to forming an interaction with DNA. b The orientation adopted by the hORC2-WHD (mauve) upon hCDC6 binding falls within the vicinity of the hMCM3-WHD (light blue) in the hOCCM structure. c The ___location of relevant COSMIC55 mutations (red) in hCDC6 (grey) in the hOCCM structure may locally disrupt interaction with hMCM3 via disruptions to inter- and intra-protein interactions. d Purified hCDC6WT, hCDC6F375C and hCDC6C394F. e Pre-RC assay under low and high salt wash conditions was carried out with hCDC6WT, hCDC6F375C and hCDC6C394F. SDS-PAGE gel is representative of four independent biological replicates. f Corresponding western blot analysis for hCDC6 (anti-hCDC6 mouse monoclonal, Santa Cruz, Cat# sc-9964) and hCDT1 (anti-hCDT1 mouse monoclonal, Santa Cruz, Cat# sc-365305) in the presence of hCDC6WT, hCDC6F375C and hCDC6C394F. ATPγS arrests complex formation at the hOCCM stage and shows maximal hCDC6 and hCDT1 association. Western blots are representative of two independent biological replicates. Source data are provided as a Source Data file.
