The SMC5 /6 complex recruits the PAF1 complex to facilitate DNA double‐strand break repair in  Arabidopsis

Abstract

DNA double‐strand breaks (DSBs) are one of the most toxic forms of DNA damage, which threatens genome stability. Homologous recombination is an error‐free DSB repair pathway, in which the evolutionarily conserved SMC5/6 complex (SMC5/6) plays essential roles. The PAF1 complex (PAF1C) is well known to regulate transcription. Here we show that SMC5/6 recruits PAF1C to facilitate DSB repair in plants. In a genetic screen for DNA damage response mutants (DDRMs), we found that the Arabidopsis ddrm4 mutant is hypersensitive to DSB‐inducing agents and is defective in homologous recombination. DDRM4 encodes PAF1, a core subunit of PAF1C. Further biochemical and genetic studies reveal that SMC5/6 recruits PAF1C to DSB sites, where PAF1C further recruits the E2 ubiquitin‐conjugating enzymes UBC1/2, which interact with the E3 ubiquitin ligases HUB1/2 to mediate the monoubiquitination of histone H2B at DSBs. These results implicate SMC5/6‐PAF1C‐UBC1/2‐HUB1/2 as a new axis for DSB repair through homologous recombination, revealing a new mechanism of SMC5/6 and uncovering a novel function of PAF1C. image The mechanisms underlying DNA double‐strand break (DSB) repair in plants are not well‐understood. In this study, the SMC5/6 complex is shown to recruit the PAF1 complex (PAF1C) to mediate H2B monoubiquitination at DSB sites, facilitating DSB repair through homologous recombination. The SMC5/6 complex recruits PAF1C to DSB sites. PAF1C recruits the E2 ubiquitin‐conjugating enzymes UBC1/2 to DSB sites. UBC1/2 recruits the E3 ubiquitin ligases HUB1/2 to DSB sites to mediate H2B monoubiquitination. PAF1C, HUB1/2, and UBC1/2 are required for DSB repair through homologous recombination.