Crystal structure of the left-handed archaeal RadA helical filament: identification of a functional motif for controlling quaternary structures and enzymatic functions of RecA family proteins

Nucleic Acids Res. 2007;35(6):1787-801. doi: 10.1093/nar/gkl1131. Epub 2007 Feb 28.

Abstract

The RecA family of proteins mediates homologous recombination, an evolutionarily conserved pathway that maintains genomic stability by protecting against DNA double strand breaks. RecA proteins are thought to facilitate DNA strand exchange reactions as closed-rings or as right-handed helical filaments. Here, we report the crystal structure of a left-handed Sulfolobus solfataricus RadA helical filament. Each protomer in this left-handed filament is linked to its neighbour via interactions of a beta-strand polymerization motif with the neighbouring ATPase domain. Immediately following the polymerization motif, we identified an evolutionarily conserved hinge region (a subunit rotation motif) in which a 360 degrees clockwise axial rotation accompanies stepwise structural transitions from a closed ring to the AMP-PNP right-handed filament, then to an overwound right-handed filament and finally to the left-handed filament. Additional structural and functional analyses of wild-type and mutant proteins confirmed that the subunit rotation motif is crucial for enzymatic functions of RecA family proteins. These observations support the hypothesis that RecA family protein filaments may function as rotary motors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / chemistry
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Archaeal Proteins / chemistry*
  • Archaeal Proteins / metabolism
  • Archaeal Proteins / ultrastructure
  • Crystallography, X-Ray
  • DNA / chemistry
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / metabolism
  • DNA-Binding Proteins / ultrastructure
  • Microscopy, Atomic Force
  • Models, Molecular*
  • Molecular Sequence Data
  • Point Mutation
  • Protein Structure, Quaternary
  • Protein Subunits / chemistry
  • Rad51 Recombinase / chemistry
  • Rec A Recombinases / chemistry*
  • Rotation
  • Saccharomyces cerevisiae Proteins / chemistry
  • Sequence Alignment
  • Sulfolobus solfataricus / enzymology

Substances

  • Archaeal Proteins
  • DNA-Binding Proteins
  • Protein Subunits
  • RadA protein, archaeal
  • Saccharomyces cerevisiae Proteins
  • Adenosine Triphosphate
  • DNA
  • RAD51 protein, S cerevisiae
  • Rad51 Recombinase
  • Rec A Recombinases