title: 53BP1 and USP28 mediate p53 activation and G1 arrest after centrosome loss or extended mitotic duration.
abstract: In normal human cells, centrosome loss induced by centrinone-a specific centrosome duplication inhibitor-leads to irreversible, p53-dependent G1 arrest by an unknown mechanism. A genome-wide CRISPR/Cas9 screen for centrinone resistance identified genes encoding the p53-binding protein 53BP1, the deubiquitinase USP28, and the ubiquitin ligase TRIM37. Deletion of TP53BP1, USP28, or TRIM37 prevented p53 elevation in response to centrosome loss but did not affect cytokinesis failure-induced arrest or p53 elevation after doxorubicin-induced DNA damage. Deletion of TP53BP1 and USP28, but not TRIM37, prevented growth arrest in response to prolonged mitotic duration. TRIM37 knockout cells formed ectopic centrosomal-component foci that suppressed mitotic defects associated with centrosome loss. TP53BP1 and USP28 knockouts exhibited compromised proliferation after centrosome removal, suggesting that centrosome-independent proliferation is not conferred solely by the inability to sense centrosome loss. Thus, analysis of centrinone resistance identified a 53BP1-USP28 module as critical for communicating mitotic challenges to the p53 circuit and TRIM37 as an enforcer of the singularity of centrosome assembly.
authors.affiliation: Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 Ludwig Institute for Cancer Research, La Jolla, CA 92093.
authors.forename: Franz
authors.identifier: 0000-0002-8635-9078
authors.initials: F
authors.lastname: Meitinger
authors.affiliation: Small Molecule Discovery Program, Ludwig Institute for Cancer Research, La Jolla, CA 92093.
authors.forename: John V
authors.identifier: 0000-0002-5376-6179
authors.initials: JV
authors.lastname: Anzola
authors.affiliation: Institute of Biochemistry II, Goethe University Frankfurt, 60590 Frankfurt am Main, Germany.
authors.forename: Manuel
authors.identifier: 0000-0002-9528-8822
authors.initials: M
authors.lastname: Kaulich
authors.affiliation: Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 Ludwig Institute for Cancer Research, La Jolla, CA 92093.
authors.forename: Amelia
authors.identifier: 0000-0002-9953-9182
authors.initials: A
authors.lastname: Richardson
authors.affiliation: Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093.
authors.forename: Joshua D
authors.identifier: 0000-0002-2263-4595
authors.initials: JD
authors.lastname: Stender
authors.affiliation: Department of Medicine, University of California, San Diego, La Jolla, California 92093.
authors.forename: Christopher
authors.initials: C
authors.lastname: Benner
authors.affiliation: Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 Department of Medicine, University of California, San Diego, La Jolla, California 92093.
authors.forename: Christopher K
authors.identifier: 0000-0003-4344-3592
authors.initials: CK
authors.lastname: Glass
authors.affiliation: Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093.
authors.forename: Steven F
authors.initials: SF
authors.lastname: Dowdy
authors.affiliation: Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 Ludwig Institute for Cancer Research, La Jolla, CA 92093.
authors.forename: Arshad
authors.initials: A
authors.lastname: Desai
authors.affiliation: Small Molecule Discovery Program, Ludwig Institute for Cancer Research, La Jolla, CA 92093.
authors.forename: Andrew K
authors.initials: AK
authors.lastname: Shiau
authors.affiliation: Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, CA 92093 Ludwig Institute for Cancer Research, La Jolla, CA 92093 koegema@ucsd.edu.
authors.forename: Karen
authors.initials: K
authors.lastname: Oegema
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D064113:CRISPR-Cas Systems / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D018385:Centrosome / Q000187:drug effects / Q000378:metabolism*; D059585:G1 Phase Cell Cycle Checkpoints / Q000187:drug effects; D017353:Gene Deletion; D055786:Gene Knockout Techniques; D005820:Genetic Testing; D006801:Humans; D008938:Mitosis / Q000187:drug effects; D009154:Mutation / Q000235:genetics; D009687:Nuclear Proteins / Q000378:metabolism; D011743:Pyrimidines / Q000494:pharmacology; D013450:Sulfones / Q000494:pharmacology; D000070584:Tripartite Motif Proteins; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D000071857:Tumor Suppressor p53-Binding Protein 1 / Q000378:metabolism*; D043222:Ubiquitin Thiolesterase / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases
chemical_list: D015415:Biomarkers; D009687:Nuclear Proteins; D011743:Pyrimidines; D013450:Sulfones; C498301:TP53BP1 protein, human; D000070584:Tripartite Motif Proteins; D016159:Tumor Suppressor Protein p53; D000071857:Tumor Suppressor p53-Binding Protein 1; C442902:USP28 protein, human; C000599097:centrinone; C411745:TRIM37 protein, human; D044767:Ubiquitin-Protein Ligases; D043222:Ubiquitin Thiolesterase