TY - JOUR
T1 - Slk19-dependent mid-anaphase pause in kinesin-5-mutated cells
AU - Movshovich, Natalia
AU - Fridman, Vladimir
AU - Gerson-Gurwitz, Adina
AU - Shumacher, Inbal
AU - Gertsberg, Irena
AU - Fich, Alexander
AU - Hoyt, M. Andrew
AU - Katz, Benjamin
AU - Gheber, Larisa
PY - 2008/8/1
Y1 - 2008/8/1
N2 - We examined spindle elongation in anaphase in Saccharomyces cerevisiae cells mutated for the kinesin-5 motor proteins Cin8 and Kip1. Cells were deleted for KIP1 and/or expressed one of two motor-domain Cin8 mutants (Cin8-F467A or Cin8-R196K, which differ in their ability to bind microtubules in vitro, with Cin8-F467A having the weakest ability). We found that, in kinesin-5-mutated cells, predominantly in kip1Δcin8-F467A cells, anaphase spindle elongation was frequently interrupted after the fast phase, resulting in a mid-anaphase pause. Expression of kinesin-5 mutants also caused an asymmetric midzone location and enlarged midzone size, suggesting that proper organization of the midzone is required for continuous spindle elongation. We also examined the effects of components of the FEAR pathway, which is involved in the early-anaphase activation of Cdc14 regulatory phosphatase, on anaphase spindle elongation in kip1Δcin8-F467A cells. Deletion of SLK19, but not SP012, eliminated the mid-anaphase pause, caused premature anaphase onset and defects in DNA division during anaphase, and reduced viability in these cells. Finally, overriding of the pre-anaphase checkpoint by overexpression of Cdc20 also eliminated the mid-anaphase pause and caused DNA deformation during anaphase in kip1Δcin8-F467A cells. We propose that transient activation of the pre-anaphase checkpoint in kinesin-5-mutated cells induces a SIk19-dependent mid-anaphase pause, which might be important for proper DNA segregation.
AB - We examined spindle elongation in anaphase in Saccharomyces cerevisiae cells mutated for the kinesin-5 motor proteins Cin8 and Kip1. Cells were deleted for KIP1 and/or expressed one of two motor-domain Cin8 mutants (Cin8-F467A or Cin8-R196K, which differ in their ability to bind microtubules in vitro, with Cin8-F467A having the weakest ability). We found that, in kinesin-5-mutated cells, predominantly in kip1Δcin8-F467A cells, anaphase spindle elongation was frequently interrupted after the fast phase, resulting in a mid-anaphase pause. Expression of kinesin-5 mutants also caused an asymmetric midzone location and enlarged midzone size, suggesting that proper organization of the midzone is required for continuous spindle elongation. We also examined the effects of components of the FEAR pathway, which is involved in the early-anaphase activation of Cdc14 regulatory phosphatase, on anaphase spindle elongation in kip1Δcin8-F467A cells. Deletion of SLK19, but not SP012, eliminated the mid-anaphase pause, caused premature anaphase onset and defects in DNA division during anaphase, and reduced viability in these cells. Finally, overriding of the pre-anaphase checkpoint by overexpression of Cdc20 also eliminated the mid-anaphase pause and caused DNA deformation during anaphase in kip1Δcin8-F467A cells. We propose that transient activation of the pre-anaphase checkpoint in kinesin-5-mutated cells induces a SIk19-dependent mid-anaphase pause, which might be important for proper DNA segregation.
KW - Anaphase B
KW - Kinesin-5
KW - Midzone
KW - Slk19
KW - Spo12
UR - http://www.scopus.com/inward/record.url?scp=50249143480&partnerID=8YFLogxK
U2 - 10.1242/jcs.022996
DO - 10.1242/jcs.022996
M3 - Article
C2 - 18628309
AN - SCOPUS:50249143480
SN - 0021-9533
VL - 121
SP - 2529
EP - 2539
JO - Journal of Cell Science
JF - Journal of Cell Science
IS - 15
ER -