Quaternary structure of the human Cdt1-Geminin complex regulates DNA replication licensing

Proc Natl Acad Sci U S A. 2009 Nov 24;106(47):19807-12. doi: 10.1073/pnas.0905281106. Epub 2009 Nov 11.

Abstract

All organisms need to ensure that no DNA segments are rereplicated in a single cell cycle. Eukaryotes achieve this through a process called origin licensing, which involves tight spatiotemporal control of the assembly of prereplicative complexes (pre-RCs) onto chromatin. Cdt1 is a key component and crucial regulator of pre-RC assembly. In higher eukaryotes, timely inhibition of Cdt1 by Geminin is essential to prevent DNA rereplication. Here, we address the mechanism of DNA licensing inhibition by Geminin, by combining X-ray crystallography, small-angle X-ray scattering, and functional studies in Xenopus and mammalian cells. Our findings show that the Cdt1:Geminin complex can exist in two distinct forms, a "permissive" heterotrimer and an "inhibitory" heterohexamer. Specific Cdt1 residues, buried in the heterohexamer, are important for licensing. We postulate that the transition between the heterotrimer and the heterohexamer represents a molecular switch between licensing-competent and licensing-defective states.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Cycle / physiology
  • Cell Cycle Proteins / chemistry*
  • Cell Cycle Proteins / genetics
  • Cell Line
  • Crystallography, X-Ray
  • DNA Replication*
  • Geminin
  • Humans
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Mutation
  • Protein Structure, Quaternary*
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Scattering, Small Angle
  • Sequence Alignment
  • X-Ray Diffraction
  • Xenopus laevis

Substances

  • CDT1 protein, human
  • Cell Cycle Proteins
  • GMNN protein, human
  • Geminin
  • Recombinant Fusion Proteins

Associated data

  • PDB/2WVR