Structural studies of the archaeal MCM complex in different functional states

J Struct Biol. 2006 Oct;156(1):210-9. doi: 10.1016/j.jsb.2006.04.001. Epub 2006 May 2.

Abstract

The primary candidate for the eukaryotic replicative helicase is the MCM2-7 complex, a hetero-oligomer formed by six AAA+ paralogous polypeptides. A simplified model for structure-function studies is the homo-oligomeric orthologue from the archaeon Methanothermobacter thermoautotrophicus. The crystal structure of the DNA-interacting N-terminal domain of this homo-oligomer revealed a double hexamer in a head-to-head configuration; single-particle electron microscopy studies have shown that the full-length protein complex can form both single and double rings, in which each ring can consist of a cyclical arrangement of six or seven subunits. Using single-particle techniques and especially multivariate statistical symmetry analysis, we have assessed the changes in stoichiometry that the complex undergoes when treated with various nucleotide analogues or when binding a double-stranded DNA fragment. We found that the binding of nucleotides or of double-stranded DNA leads to the preferred formation of double-ring structures. Specifically, the protein complex is present as a double heptamer when treated with a nucleotide analogue, but it is rather found as a double hexamer when complexed with double-stranded DNA. The possible physiological role of the various stoichiometries of the complex is discussed in the light of the proposed mechanisms of helicase activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Diphosphate / pharmacology
  • Archaeal Proteins / chemistry
  • Archaeal Proteins / isolation & purification
  • Archaeal Proteins / metabolism*
  • Archaeal Proteins / ultrastructure
  • Chromosomes, Archaeal / chemistry
  • Chromosomes, Archaeal / metabolism*
  • DNA / metabolism
  • DNA Helicases / chemistry*
  • DNA Helicases / genetics
  • DNA Helicases / metabolism*
  • Escherichia coli / genetics
  • Methanobacteriaceae / enzymology
  • Models, Biological
  • Nucleotides / metabolism
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Tertiary
  • Substrate Specificity
  • Transformation, Genetic

Substances

  • Archaeal Proteins
  • Nucleotides
  • Adenosine Diphosphate
  • DNA
  • DNA Helicases