TY - JOUR
T1 - DIDS, a chemical compound that inhibits RAD51-mediated homologous pairing and strand exchange
AU - Ishida, Takako
AU - Takizawa, Yoshimasa
AU - Kainuma, Takashi
AU - Inoue, Jin
AU - Mikawa, Tsutomu
AU - Shibata, Takehiko
AU - Suzuki, Hidekazu
AU - Tashiro, Satoshi
AU - Kurumizaka, Hitoshi
N1 - Funding Information:
Ministry of Education, Culture, Sports, Science and Technology, Japan; Chemical compounds: Scientific Research on Priority Areas, Cancer, Japan. Funding for open access charge: Waseda University.
PY - 2009
Y1 - 2009
N2 - RAD51, an essential eukaryotic DNA recombinase, promotes homologous pairing and strand exchange during homologous recombination and the recombinational repair of double strand breaks. Mutations that up- or down-regulate RAD51 gene expression have been identified in several tumors, suggesting that inappropriate expression of the RAD51 activity may cause tumorigenesis. To identify chemical compounds that affect the RAD51 activity, in the present study, we performed the RAD51-mediated strand exchange assay in the presence of 185 chemical compounds. We found that 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (DIDS) efficiently inhibited the RAD51-mediated strand exchange. DIDS also inhibited the RAD51-mediated homologous pairing in the absence of RPA. A surface plasmon resonance analysis revealed that DIDS directly binds to RAD51. A gel mobility shift assay showed that DIDS significantly inhibited the DNA-binding activity of RAD51. Therefore, DIDS may bind near the DNA binding site(s) of RAD51 and compete with DNA for RAD51 binding.
AB - RAD51, an essential eukaryotic DNA recombinase, promotes homologous pairing and strand exchange during homologous recombination and the recombinational repair of double strand breaks. Mutations that up- or down-regulate RAD51 gene expression have been identified in several tumors, suggesting that inappropriate expression of the RAD51 activity may cause tumorigenesis. To identify chemical compounds that affect the RAD51 activity, in the present study, we performed the RAD51-mediated strand exchange assay in the presence of 185 chemical compounds. We found that 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (DIDS) efficiently inhibited the RAD51-mediated strand exchange. DIDS also inhibited the RAD51-mediated homologous pairing in the absence of RPA. A surface plasmon resonance analysis revealed that DIDS directly binds to RAD51. A gel mobility shift assay showed that DIDS significantly inhibited the DNA-binding activity of RAD51. Therefore, DIDS may bind near the DNA binding site(s) of RAD51 and compete with DNA for RAD51 binding.
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U2 - 10.1093/nar/gkp200
DO - 10.1093/nar/gkp200
M3 - Article
C2 - 19336413
AN - SCOPUS:67249134799
SN - 0305-1048
VL - 37
SP - 3367
EP - 3376
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 10
ER -