Rad51 Interaction Analysis Reveals a Functional Interplay Among Recombination Auxiliary Factors
Rad51 Interaction Analysis Reveals a Functional Interplay Among Recombination Auxiliary Factors
ABSTRACT Although Rad51 is the key protein in homologous recombination (HR), a major DNA double-strand break repair pathway, several auxiliary factors interact with Rad51 to promote productive HR. Here, we present an interdisciplinary characterization of the interaction between Rad51 and Swi5-Sfr1, a widely conserved auxiliary factor. NMR and site-specific crosslinking experiments revealed two distinct sites within the intrinsically disordered N-terminus of Sfr1 that cooperatively bind to Rad51. Although disruption of this binding severely impaired Rad51 stimulation in vitro, interaction mutants did not show any defects in DNA repair. Unexpectedly, in the absence of the Rad51 paralogs Rad55-Rad57, which constitute another auxiliary factor complex, these interaction mutants were unable to promote DNA repair. Our findings provide molecular insights into Rad51 stimulation by Swi5-Sfr1 and suggest that, rather than functioning in an independent subpathway of HR as was previously proposed, Rad55-Rad57 facilitates the recruitment of Swi5-Sfr1 to Rad51.
Sakakura Masayoshi、Kurihara Misato、Murayama Yasuto、Takahashi Masayuki、Takahashi Hideo、Argunhan Bilge、Maki Takahisa、Tsubouchi Hideo、Iwasaki Hiroshi、Kanamaru Shuji、Ito Kentaro、Afshar Negar
Graduate School of Medical Life Science, Yokohama City UniversityGraduate School of Medical Life Science, Yokohama City University||Nomura Research InstituteCenter for Frontier Research, National Institute of GeneticsSchool of Life Science and Technology, Tokyo Institute of TechnologyGraduate School of Medical Life Science, Yokohama City UniversityInstitute of Innovative Research, Tokyo Institute of TechnologyInstitute of Innovative Research, Tokyo Institute of TechnologyInstitute of Innovative Research, Tokyo Institute of TechnologyInstitute of Innovative Research, Tokyo Institute of Technology||School of Life Science and Technology, Tokyo Institute of TechnologyInstitute of Innovative Research, Tokyo Institute of TechnologyInstitute of Innovative Research, Tokyo Institute of TechnologyInstitute of Innovative Research, Tokyo Institute of Technology
分子生物学遗传学细胞生物学
Homologous recombinationRad51auxiliary factorintrinsically disorderedSwi5-Sfr1Rad55-Rad57Rad51 paralogsprotein NMR
Sakakura Masayoshi,Kurihara Misato,Murayama Yasuto,Takahashi Masayuki,Takahashi Hideo,Argunhan Bilge,Maki Takahisa,Tsubouchi Hideo,Iwasaki Hiroshi,Kanamaru Shuji,Ito Kentaro,Afshar Negar.Rad51 Interaction Analysis Reveals a Functional Interplay Among Recombination Auxiliary Factors[EB/OL].(2025-03-28)[2025-06-07].https://www.biorxiv.org/content/10.1101/738179.点此复制
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