Centrobin regulates the assembly of functional mitotic spindles.
The proper function of the spindle is crucial to the high fidelity of chromosome segregation and is indispensable for tumor suppression in humans. Centrobin is a recently identified centrosomal protein that has a role in stabilizing the microtubule structure. Here we functionally characterize the defects in centrosome integrity and spindle assembly in Centrobin-depleted cells. Centrobin-depleted cells show a range of spindle abnormalities including unfocused poles that are not associated with centrosomes, S-shaped spindles and mini spindles. These cells undergo mitotic arrest and subsequently often die by apoptosis, as determined by live cell imaging. Co-depletion of Mad2 relieves the mitotic arrest, indicating that cells arrest due to a failure to silence the spindle checkpoint in metaphase. Consistent with this, Centrobin-depleted metaphase cells stained positive for BubR1 and BubR1 S676. Staining with a panel of centrosome markers showed a loss of centrosome anchoring to the mitotic spindle. Furthermore, these cells show less cold-stable microtubules and a shorter distance between kinetochore pairs. These results show a requirement of Centrobin in maintaining centrosome integrity, which in turn promotes anchoring of mitotic spindle to the centrosomes. Furthermore, this anchoring is required for the stability of microtubule-kinetochore attachments and biogenesis of tension-ridden and properly functioning mitotic spindle.
Jeffery
J M
JM
Signal Transduction Laboratory, Cancer and Cell Biology Division, Queensland Institute of Medical Research, Brisbane, Queensland, Australia.
Urquhart
A J
AJ
Subramaniam
V N
VN
Parton
R G
RG
Khanna
K K
KK
D034961:Antigens, Nuclear / Q000032:analysis; D002135:Calcium-Binding Proteins / Q000502:physiology; D018797:Cell Cycle Proteins / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D064246:Mad2 Proteins; D000072379:NIMA-Related Kinases; D034521:Nuclear Matrix-Associated Proteins / Q000032:analysis; D017346:Protein Serine-Threonine Kinases / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000502:physiology; D012097:Repressor Proteins / Q000502:physiology; D008941:Spindle Apparatus / Q000502:physiology*; D014404:Tubulin / Q000032:analysis; D000097869:Polo-Like Kinase 1
D034961:Antigens, Nuclear; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D064246:Mad2 Proteins; D000072379:NIMA-Related Kinases; D034521:Nuclear Matrix-Associated Proteins; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D012097:Repressor Proteins; D014404:Tubulin; D000097869:Polo-Like Kinase 1; C507203:CNTROB protein, human; C577697:MAD2L1 protein, human; C050045:NUMA1 protein, human; C089441:NEK2 protein, human