The ESCRT machinery counteracts Nesprin-2G-mediated mechanical forces during nuclear envelope repair

TY - JOUR

T1 - The ESCRT machinery counteracts Nesprin-2G-mediated mechanical forces during nuclear envelope repair

AU - Wallis, Samuel S

AU - Ventimiglia, Leandro N

AU - Otigbah, Evita

AU - Infante, Elvira

AU - Cuesta-Geijo, Miguel Angel

AU - Kidiyoor, Gururaj Rao

AU - Carbajal, M Alejandra

AU - Fleck, Roland A

AU - Foiani, Marco

AU - Garcia-Manyes, Sergi

AU - Martin-Serrano, Juan

AU - Agromayor, Monica

N1 - Funding Information: The authors thank W. H. DeVos for kindly providing the lamina/C knockout cells, the UK NIHR Comprehensive BRC at KCL for an equipment grant, and the Nikon Imaging Centre at KCL for technical support in imaging. This work was supported by awards from the BBSRC (BB/N000501/1) and the King's Health Partners R&D research fund (R151001) to M.A.; the Wellcome Trust (WT102871MA) to J.M.-S.; the European Commission (FET Proactive 731957), EPSRC (EP/M022536/1), Leverhulme Trust (RPG-2015-225 and RL-2016-015), Wellcome Trust (212218/Z/18/Z) and Royal Society (RSWF/R3/183006) to S.G.-M.; and the Italian Association for Cancer Research (AIRC) to M.F. S.S.W. was supported by the King's Bioscience Institute and the Guy's and St Thomas? Charity Prize Ph.D. Programme in Biomedical and Translational Science. E.O. is supported by the UK Medical Research Council (MR/N013700/1) and is a King's College London member of the MRC Doctoral Training Partnership in Biomedical Sciences. S.S.W. L.N.V. E.O. J.M.-S. and M.A. conceived the study. E.I. and S.G.-M. designed, performed, and analyzed AFM experiments. M.A.C. and R.A.F. designed, performed, and analyzed electron microscopy experiments. S.S.W. L.N.V. E.O. and M.A.C.-G. generated tools and performed and analyzed data from all other experiments. G.R.K. and M.F. helped with the design and analysis of experiments using microchannels. M.A. and J.M.-S. analyzed data and wrote the manuscript with assistance from all other authors. The authors declare no competing interests. Funding Information: The authors thank W. H. DeVos for kindly providing the lamina/C knockout cells, the UK NIHR Comprehensive BRC at KCL for an equipment grant, and the Nikon Imaging Centre at KCL for technical support in imaging. This work was supported by awards from the BBSRC ( BB/N000501/1 ) and the King’s Health Partners R&D research fund ( R151001 ) to M.A.; the Wellcome Trust ( WT102871MA ) to J.M.-S.; the European Commission (FET Proactive 731957), EPSRC ( EP/M022536/1 ), Leverhulme Trust ( RPG-2015-225 and RL-2016-015 ), Wellcome Trust ( 212218/Z/18/Z ) and Royal Society ( RSWF/R3/183006 ) to S.G.-M.; and the Italian Association for Cancer Research (AIRC) to M.F. S.S.W. was supported by the King’s Bioscience Institute and the Guy’s and St Thomas’ Charity Prize Ph.D. Programme in Biomedical and Translational Science . E.O. is supported by the UK Medical Research Council ( MR/N013700/1 ) and is a King’s College London member of the MRC Doctoral Training Partnership in Biomedical Sciences . Publisher Copyright: © 2021 The Authors

PY - 2021/12/6

Y1 - 2021/12/6

N2 - Transient nuclear envelope ruptures during interphase (NERDI) occur due to cytoskeletal compressive forces at sites of weakened lamina, and delayed NERDI repair results in genomic instability. Nuclear envelope (NE) sealing is completed by endosomal sorting complex required for transport (ESCRT) machinery. A key unanswered question is how local compressive forces are counteracted to allow efficient membrane resealing. Here, we identify the ESCRT-associated protein BROX as a crucial factor required to accelerate repair of the NE. Critically, BROX binds Nesprin-2G, a component of the linker of nucleoskeleton and cytoskeleton complex (LINC). This interaction promotes Nesprin-2G ubiquitination and facilitates the relaxation of mechanical stress imposed by compressive actin fibers at the rupture site. Thus, BROX rebalances excessive cytoskeletal forces in cells experiencing NE instability to promote effective NERDI repair. Our results demonstrate that BROX coordinates mechanoregulation with membrane remodeling to ensure the maintenance of nuclear-cytoplasmic compartmentalization and genomic stability.

AB - Transient nuclear envelope ruptures during interphase (NERDI) occur due to cytoskeletal compressive forces at sites of weakened lamina, and delayed NERDI repair results in genomic instability. Nuclear envelope (NE) sealing is completed by endosomal sorting complex required for transport (ESCRT) machinery. A key unanswered question is how local compressive forces are counteracted to allow efficient membrane resealing. Here, we identify the ESCRT-associated protein BROX as a crucial factor required to accelerate repair of the NE. Critically, BROX binds Nesprin-2G, a component of the linker of nucleoskeleton and cytoskeleton complex (LINC). This interaction promotes Nesprin-2G ubiquitination and facilitates the relaxation of mechanical stress imposed by compressive actin fibers at the rupture site. Thus, BROX rebalances excessive cytoskeletal forces in cells experiencing NE instability to promote effective NERDI repair. Our results demonstrate that BROX coordinates mechanoregulation with membrane remodeling to ensure the maintenance of nuclear-cytoplasmic compartmentalization and genomic stability.

UR - http://www.scopus.com/inward/record.url?scp=85120169353&partnerID=8YFLogxK

U2 - 10.1016/j.devcel.2021.10.022

DO - 10.1016/j.devcel.2021.10.022

M3 - Article

C2 - 34818527

SN - 1534-5807

VL - 56

SP - 3192-3202.e8

JO - Developmental Cell

JF - Developmental Cell

IS - 23

ER -