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PARP1-DNA co-condensation drives DNA repair site assembly to prevent disjunction of broken DNA ends

Cell. 2024-01; 
Nagaraja Chappidi 1, Thomas Quail 2, Simon Doll 3, Laura T Vogel 4, Radoslav Aleksandrov 5, Suren Felekyan 4, Ralf Kühnemuth 4, Stoyno Stoynov 5, Claus A M Seidel 4, Jan Brugués 6, Marcus Jahnel 3, Titus M Franzmann 1, Simon Alberti
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Codon Optimization PARP1, PARP2, PARG, HPF1, FUS, EWSR1 and TAF15 constructs were synthesized (GenScript) as codon-optimized genes for expression in Spodoptera frugiperda Sf9 cells. Get A Quote
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摘要

DNA double-strand breaks (DSBs) are repaired at DSB sites. How DSB sites assemble and how broken DNA is prevented from separating is not understood. Here we uncover that the synapsis of broken DNA is mediated by the DSB sensor protein poly(ADP-ribose) (PAR) polymerase 1 (PARP1). Using bottom-up biochemistry, we reconstitute functional DSB sites and show that DSB sites form through co-condensation of PARP1 multimers with DNA. The co-condensates exert mechanical forces to keep DNA ends together and become enzymatically active for PAR synthesis. PARylation promotes release of PARP1 from DNA ends and the recruitment of effectors, such as Fused in Sarcoma, which stabilizes broken DNA ends against separation, reveali... More

关键词

DNA damage repair; DNA double-strand break; DNA end synapsis; FET proteins; PARP1; PARylation; condensate; dimerization; multimerization; phase separation.