53BP1 and USP28 mediate p53 activation and G1 arrest after centrosome loss or extended mitotic duration

J Cell Biol. 2016 Jul 18;214(2):155-66. doi: 10.1083/jcb.201604081.

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.

MeSH terms

  • Biomarkers / metabolism
  • CRISPR-Cas Systems / genetics
  • Cell Line
  • Cell Proliferation / drug effects
  • Centrosome / drug effects
  • Centrosome / metabolism*
  • G1 Phase Cell Cycle Checkpoints* / drug effects
  • Gene Deletion
  • Gene Knockout Techniques
  • Genetic Testing
  • Humans
  • Mitosis* / drug effects
  • Mutation / genetics
  • Nuclear Proteins / metabolism
  • Pyrimidines / pharmacology
  • Sulfones / pharmacology
  • Tripartite Motif Proteins
  • Tumor Suppressor Protein p53 / metabolism*
  • Tumor Suppressor p53-Binding Protein 1 / metabolism*
  • Ubiquitin Thiolesterase / metabolism*
  • Ubiquitin-Protein Ligases

Substances

  • Biomarkers
  • Nuclear Proteins
  • Pyrimidines
  • Sulfones
  • TP53BP1 protein, human
  • Tripartite Motif Proteins
  • Tumor Suppressor Protein p53
  • Tumor Suppressor p53-Binding Protein 1
  • USP28 protein, human
  • centrinone
  • TRIM37 protein, human
  • Ubiquitin-Protein Ligases
  • Ubiquitin Thiolesterase