Biochemical analysis of the human EVL domains in homologous recombination.
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).
Takaku
Motoki
M
Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering, Waseda University, Tokyo, Japan.
Machida
Shinichi
S
Nakayama
Shugo
S
Takahashi
Daisuke
D
Kurumizaka
Hitoshi
H
D015815:Cell Adhesion Molecules / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding / Q000235:genetics / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000235:genetics / Q000502:physiology; D051135:Rad51 Recombinase / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics / Q000502:physiology*
D015815:Cell Adhesion Molecules; C484449:EVL protein, human; C495153:RAD51 protein, human; D051135:Rad51 Recombinase