RNA-binding proteins (RBPs) are emerging as important effectors of the cellular DNA damage response (DDR). The RBP FUS is implicated in RNA metabolism and DNA repair, and it undergoes reversible liquid-liquid phase separation (LLPS) in vitro. Here, we demonstrate that FUS-dependent LLPS is necessary for the initiation of the DDR. Using laser microirradiation in FUS-knockout cells, we show that FUS is required for the recruitment to DNA damage sites of the DDR factors KU80, NBS1, 53BP1, and of SFPQ, another RBP implicated in the DDR. The relocation of KU80, NBS1, and SFPQ is similarly impaired by LLPS inhibitors, or LLPS-deficient FUS variants. We also show that LLPS is necessary for efficient γH2AX foci formation. Finally, using super-resolution structured illumination microscopy, we demonstrate that the absence of FUS impairs the proper arrangement of γH2AX nano-foci into higher-order clusters. These findings demonstrate the early requirement for FUS-dependent LLPS in the activation of the DDR and the proper assembly of DSBs repair complexes.

Rocha Levone, B., Lenzken, S., Antonaci, M., Maiser, A., Rapp, A., Conte, F., et al. (2019). FUS-dependent liquid-liquid phase separation is an early event in double-strand break repair [Altro].

FUS-dependent liquid-liquid phase separation is an early event in double-strand break repair

Rocha Levone, B
Primo
Membro del Collaboration Group
;
Lenzken, SC;Ronchi, AE;Barabino, SML
Ultimo
Membro del Collaboration Group
2019

Abstract

RNA-binding proteins (RBPs) are emerging as important effectors of the cellular DNA damage response (DDR). The RBP FUS is implicated in RNA metabolism and DNA repair, and it undergoes reversible liquid-liquid phase separation (LLPS) in vitro. Here, we demonstrate that FUS-dependent LLPS is necessary for the initiation of the DDR. Using laser microirradiation in FUS-knockout cells, we show that FUS is required for the recruitment to DNA damage sites of the DDR factors KU80, NBS1, 53BP1, and of SFPQ, another RBP implicated in the DDR. The relocation of KU80, NBS1, and SFPQ is similarly impaired by LLPS inhibitors, or LLPS-deficient FUS variants. We also show that LLPS is necessary for efficient γH2AX foci formation. Finally, using super-resolution structured illumination microscopy, we demonstrate that the absence of FUS impairs the proper arrangement of γH2AX nano-foci into higher-order clusters. These findings demonstrate the early requirement for FUS-dependent LLPS in the activation of the DDR and the proper assembly of DSBs repair complexes.
Altro
Preprint paper available online in the following link: https://www.biorxiv.org/content/10.1101/798884v2
RNA-binding proteins, FUS, SFPQ, DNA damage repair, double strand breaks, phase separation
English
2019
2019
https://www.biorxiv.org/content/10.1101/798884v2
Rocha Levone, B., Lenzken, S., Antonaci, M., Maiser, A., Rapp, A., Conte, F., et al. (2019). FUS-dependent liquid-liquid phase separation is an early event in double-strand break repair [Altro].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/276080
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