[en] Eukaryotic cells have evolved highly orchestrated protein catabolic machineries responsible for the timely and selective disposal of proteins and organelles, thereby ensuring amino acid recycling. However, how protein degradation is coordinated with amino acid supply and protein synthesis has remained largely elusive. Here we show that the mammalian proteasome undergoes liquid-liquid phase separation in the nucleus upon amino acid deprivation. We termed these proteasome condensates SIPAN (Starvation-Induced Proteasome Assemblies in the Nucleus) and show that these are a common response of mammalian cells to amino acid deprivation. SIPAN undergo fusion events, rapidly exchange proteasome particles with the surrounding milieu and quickly dissolve following amino acid replenishment. We further show that: (i) SIPAN contain K48-conjugated ubiquitin, (ii) proteasome inhibition accelerates SIPAN formation, (iii) deubiquitinase inhibition prevents SIPAN resolution and (iv) RAD23B proteasome shuttling factor is required for SIPAN formation. Finally, SIPAN formation is associated with decreased cell survival and p53-mediated apoptosis, which might contribute to tissue fitness in diverse pathophysiological conditions.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Uriarte, Maxime ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques ; Department of Biochemistry and Molecular Medicine, University of Montréal, H3C 3J7, Montreal, QC, Canada ; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada
Sen Nkwe, Nadine; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Molecular Biology Programs, University of Montreal, Montréal, H3A 0G4, QC, Canada
Tremblay, Roch; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Molecular Biology Programs, University of Montreal, Montréal, H3A 0G4, QC, Canada
Ahmed, Oumaima; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Molecular Biology Programs, University of Montreal, Montréal, H3A 0G4, QC, Canada
Messmer, Clémence; Department of Biochemistry and Molecular Medicine, University of Montréal, H3C 3J7, Montreal, QC, Canada ; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada
Mashtalir, Nazar; Department of Pediatric Oncology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA, 02215, USA ; Broad Institute of MIT and Harvard, Cambridge, MA, 02142, USA
Barbour, Haithem; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Biomedical Sciences Programs, University of Montréal, Montréal, H3C 3T5, QC, Canada
Masclef, Louis ; Department of Biochemistry and Molecular Medicine, University of Montréal, H3C 3J7, Montreal, QC, Canada ; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada
Voide, Marion; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Molecular Biology Programs, University of Montreal, Montréal, H3A 0G4, QC, Canada
Viallard, Claire ; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Department of Medicine, University of Montréal, Montréal, H3C 3J7, QC, Canada
Daou, Salima; Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON, M5G 1X5, Canada
Abdelhadi, Djaileb; Department of Biochemistry and Molecular Medicine, University of Montréal, H3C 3J7, Montreal, QC, Canada ; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada
Ronato, Daryl ; CHU de Québec Research Center, Oncology Division, 9 McMahon, Québec City, QC, G1R 3S3, Canada ; Laval University Cancer Research Center, Québec City, QC, G1V 0A6, Canada
Paydar, Mohammadjavad; Institute for Research in Immunology and Cancer (IRIC), University of Montréal, Montréal, QC, H3T 1J4, Canada
Darracq, Anaïs; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Molecular Biology Programs, University of Montreal, Montréal, H3A 0G4, QC, Canada
Boulay, Karine; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Department of Medicine, University of Montréal, Montréal, H3C 3J7, QC, Canada
Desjardins-Lecavalier, Nicolas ; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada
Sapieha, Przemyslaw ; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Department of Ophthalmology, University of Montréal, Montréal, QC, Canada ; Department of Neurology-Neurosurgery, McGill University, Montréal, QC, Canada
Masson, Jean-Yves ; CHU de Québec Research Center, Oncology Division, 9 McMahon, Québec City, QC, G1R 3S3, Canada ; Laval University Cancer Research Center, Québec City, QC, G1V 0A6, Canada
Sergeev, Mikhail; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Department of Medicine, University of Montréal, Montréal, H3C 3J7, QC, Canada
Kwok, Benjamin H; Department of Medicine, University of Montréal, Montréal, H3C 3J7, QC, Canada ; Institute for Research in Immunology and Cancer (IRIC), University of Montréal, Montréal, QC, H3T 1J4, Canada
Hulea, Laura; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Department of Medicine, University of Montréal, Montréal, H3C 3J7, QC, Canada
Mallette, Frédérick A ; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Department of Medicine, University of Montréal, Montréal, H3C 3J7, QC, Canada
Milot, Eric; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Department of Medicine, University of Montréal, Montréal, H3C 3J7, QC, Canada
Larrivée, Bruno; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Department of Medicine, University of Montréal, Montréal, H3C 3J7, QC, Canada
Wurtele, Hugo; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada ; Department of Medicine, University of Montréal, Montréal, H3C 3J7, QC, Canada
Affar, El Bachir ; Maisonneuve-Rosemont Hospital Research Center, Montréal, QC, H1T 2M4, Canada. el.bachir.affar@umontreal.ca ; Department of Medicine, University of Montréal, Montréal, H3C 3J7, QC, Canada. el.bachir.affar@umontreal.ca
CIHR - Canadian Institutes of Health Research NSERC - Natural Sciences and Engineering Research Council
Funding text :
We thank Diana Adjaoud for technical assistance, Dr. Diane Gingras for help with electronic microscopy and Dr. Alain Verreault for yeast strains. This work was supported by grants from the Canadian Institutes of Health Research (CIHR) (MOP159539) to E.B.A., CIHR (GER-163050) to K.B. and F.A.M., the Natural Sciences and Engineering Research Council of Canada (NSERC) (DG 2015–2021) and The Cancer Research Society (2015–2017) to E.B.A., NSERC (DG 2019–2024) to H.W., CIHR (MOP126009) to E.M., the CIHR (Foundation grant to J.-Y.M.). E.B.A., L.H., B.L., and H.W are scholars of the Fonds de la Recherche du Québec-Santé (FRQ-S). J.-Y.M. is a FRQS Chair in genome stability. F.A.M. holds the Canada Research Chair In Epigenetics of Aging and Cancer. K.B. is the recipient of a Cole Foundation post-doctoral fellowship. N.K. and O.A had PhD scholarships from the FRQ-S and the Cole Foundation. L.M. had a PhD scholarship from the FRQ-S.
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