The Chfr mitotic checkpoint protein functions with Ubc13-Mms2 to form Lys63-linked polyubiquitin chains.
We recently described a novel checkpoint pathway that functions early in mitosis to delay chromosome condensation in response to microtubule poisons. The only gene implicated so far in this checkpoint pathway is chfr, whose protein product contains a RING domain and has ubiquitin ligase activity in vitro. The significance of this activity in vivo is unclear. A recent report suggested that the Chfr protein targets itself for proteasome-dependent degradation in mitotic cells through autoubiquitination. However, we observe that in mitosis Chfr exhibits a phosphorylation-dependent electrophoretic mobility shift with no change in overall protein levels. Further analysis of its ubiquitin ligase activity revealed that Chfr can catalyse the formation of noncanonical Lys63-linked polyubiquitin chains with Ubc13-Mms2 acting as the ubiquitin-conjugating enzyme. Ubc13-Mms2 and Lys63-polyubiquitin chains are not associated with targeting proteins to the proteasome, but rather with signaling cellular stress. We propose that Chfr may have a role in signaling the presence of mitotic stress induced by microtubule poisons.
Bothos
John
J
The Wistar Institute, Philadelphia, PA 19104-4268, USA.
Summers
Matthew K
MK
Venere
Monica
M
Scolnick
Daniel M
DM
Halazonetis
Thanos D
TD
D002453:Cell Cycle / Q000235:genetics / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000378:metabolism / Q000502:physiology*; D003001:Cloning, Molecular; D004260:DNA Repair / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000502:physiology*; D006801:Humans; D008870:Microtubules / Q000502:physiology / Q000648:ultrastructure; D008940:Mitotic Index; D009363:Neoplasm Proteins; D012516:Osteosarcoma; D010766:Phosphorylation; D000075223:Poly-ADP-Ribose Binding Proteins; D011994:Recombinant Proteins / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D014407:Tumor Cells, Cultured; D025801:Ubiquitin / Q000378:metabolism*; D044763:Ubiquitin-Conjugating Enzymes / Q000502:physiology*; D044767:Ubiquitin-Protein Ligases
D018797:Cell Cycle Proteins; D005656:Fungal Proteins; C112520:MMS2 protein, S cerevisiae; D009363:Neoplasm Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; D044763:Ubiquitin-Conjugating Enzymes; C412002:CHFR protein, human; D044767:Ubiquitin-Protein Ligases