The DASH complex and Klp5/Klp6 kinesin coordinate bipolar chromosome attachment in fission yeast.
We identified a truncated allele of dam1 as a multicopy suppressor of the sensitivity of cdc13-117 (cyclin B) and mal3-1 (EB-1) cells to thiabendazole, a microtubule poison. We find that Dam1 binds to the plus end of spindle microtubules and kinetochores as cells enter mitosis and this is dependent on other components of the fission yeast DASH complex, including Ask1, Duo1, Spc34 and Dad1. By contrast, Dad1 remains bound to kinetochores throughout the cell cycle and its association is dependent on the Mis6 and Mal2, but not Mis12, Nuf2 or Cnp1, kinetochore proteins. In cells lacking Dam1, or other components of the DASH complex, anaphase is delayed due to activation of the spindle assembly checkpoint and lagging sister chromatids are frequently observed and occasionally sister chromatid pairs segregate to the same spindle pole. We find that the mitotic centromere-associated Klp5/Klp6 kinesin complex is essential in cells lacking components of the DASH complex. Cells lacking both Dam1 and Klp5 undergo a first cell cycle arrest in mitosis due to a failure to establish bipolar chromosome attachment.
Sanchez-Perez
Isabel
I
Division of Yeast Genetics, National Institute for Medical Research, London, UK.
Renwick
Steven J
SJ
Crawley
Karen
K
Karig
Inga
I
Buck
Vicky
V
Meadows
John C
JC
Franco-Sanchez
Alejandro
A
Fleig
Ursula
U
Toda
Takashi
T
Millar
Jonathan B A
JB
D020090:Chromosome Segregation / Q000502:physiology*; D015825:Chromosomes, Fungal / Q000378:metabolism* / Q000502:physiology; D003001:Cloning, Molecular; D019926:Cyclin B; D017931:DNA Primers; D016153:Genes, Suppressor / Q000502:physiology; D016547:Kinesins / Q000235:genetics / Q000378:metabolism*; D046911:Macromolecular Substances / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008938:Mitosis / Q000502:physiology*; D012568:Schizosaccharomyces; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*; D013827:Thiabendazole
D019926:Cyclin B; D017931:DNA Primers; D046911:Macromolecular Substances; D008869:Microtubule-Associated Proteins; D029702:Schizosaccharomyces pombe Proteins; C469734:Klp5 protein, S pombe; C459102:Klp6 protein, S pombe; D016547:Kinesins; D013827:Thiabendazole