Mid-anaphase arrest in S. cerevisiae cells eliminated for the function of Cin8 and dynein.
S. cerevisiae anaphase spindle elongation is accomplished by the overlapping function of dynein and the kinesin-5 motor proteins, Cin8 and Kip1. Cin8 and dynein are synthetically lethal, yet the arrest phenotypes of cells eliminated for their function had not been identified. We found that at a non-permissive temperature, dyn1 Delta cells that carry a temperature-sensitive cin8 - 3 mutation arrest at mid-anaphase with a unique phenotype, which we named TAN (two microtubule asters in one nucleus). These cells enter anaphase, but fail to proceed through the slow phase of anaphase B. At a permissive temperature, dyn1 Delta, cin8 - 3 or dyn1 Delta cin8 - 3 cells exhibit perturbed spindle midzone morphologies, with dyn1Delta cin8 - 3 anaphase spindles also being profoundly bent and nonrigid. Sorbitol, which has been suggested to stabilize microtubules, corrects these defects and suppresses the TAN phenotype. We conclude that dynein and Cin8 cooperate in anaphase midzone organization and influence microtubule dynamics, thus enabling progression through the slow phase of anaphase B.
Gerson-Gurwitz
A
A
Department of Chemistry, Ben-Gurion University of the Negev, Beer-Sheva, 84105, Israel.
Movshovich
N
N
Avunie
R
R
Fridman
V
V
Moyal
K
K
Katz
B
B
Hoyt
M A
MA
Gheber
L
L
D000705:Anaphase / Q000502:physiology*; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D004398:Dyneins / Q000235:genetics / Q000378:metabolism*; D007202:Indicators and Reagents / Q000378:metabolism; D016547:Kinesins; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D009154:Mutation; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013012:Sorbitol / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism / Q000648:ultrastructure
C075342:CIN8 protein, S cerevisiae; D007202:Indicators and Reagents; D008869:Microtubule-Associated Proteins; D029701:Saccharomyces cerevisiae Proteins; D013012:Sorbitol; D004398:Dyneins; D016547:Kinesins