Biochemical analysis of the human EVL domains in homologous recombination

FEBS J. 2009 Oct;276(20):5841-8. doi: 10.1111/j.1742-4658.2009.07265.x. Epub 2009 Sep 2.

Abstract

EVL is a member of the ENA/VASP family, which is involved in actin-remodeling processes. Previously, we reported that human EVL directly interacts with RAD51, which is an essential protein in the homologous recombinational repair of DNA double-strand breaks, and stimulates RAD51-mediated recombination reactions in vitro. To identify the EVL domain required for the recombination function, we purified the EVL fragments EVL(1-221) and EVL(222-418), which contain the EVH1 and Pro-rich domains and the EVH2 domain, respectively. We found that EVL(222-418) possesses DNA-binding and RAD51-binding activities, and also stimulates RAD51-mediated homologous pairing. In contrast, EVL(1-221) did not exhibit any of these activities. Therefore, the EVH2 domain, which is highly conserved among the ENA/VASP family proteins, may be responsible for the recombination function of EVL. Structured digital abstract: * MINT-7239394: EVL (uniprotkb:Q9UI08) binds (MI:0407) to RAD51 (uniprotkb:Q06609) by pull down (MI:0096).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Adhesion Molecules / chemistry*
  • Cell Adhesion Molecules / genetics
  • Cell Adhesion Molecules / metabolism*
  • Humans
  • Protein Binding / genetics
  • Protein Binding / physiology
  • Protein Structure, Tertiary / genetics
  • Protein Structure, Tertiary / physiology
  • Rad51 Recombinase / metabolism
  • Recombination, Genetic / genetics
  • Recombination, Genetic / physiology*

Substances

  • Cell Adhesion Molecules
  • EVL protein, human
  • RAD51 protein, human
  • Rad51 Recombinase