Requirement for the budding yeast polo kinase Cdc5 in proper microtubule growth and dynamics.
In many organisms, polo kinases appear to play multiple roles during M-phase progression. To provide new insights into the function of the budding yeast polo kinase Cdc5, we generated novel temperature-sensitive cdc5 mutants by mutagenizing the C-terminal noncatalytic polo box domain, a region that is critical for proper subcellular localization. One of these mutants, cdc5-11, exhibited a temperature-sensitive growth defect with an abnormal spindle morphology. Strikingly, provision of a moderate level of benomyl, a microtubule-depolymerizing drug, permitted cdc5-11 cells to grow significantly better than the isogenic CDC5 wild type in a FEAR (cdc Fourteen Early Anaphase Release)-independent manner. In addition, cdc5-11 required MAD2 for both cell growth and the benomyl-remedial phenotype. These results suggest that cdc5-11 is defective in proper spindle function. Consistent with this view, cdc5-11 exhibited abnormal spindle morphology, shorter spindle length, and delayed microtubule regrowth at the nonpermissive temperature. Overexpression of CDC5 moderately rescued the spc98-2 growth defect. Interestingly, both Cdc28 and Cdc5 were required for the proper modification of the spindle pole body components Nud1, Slk19, and Stu2 in vivo. They also phosphorylated these three proteins in vitro. Taken together, these observations suggest that concerted action of Cdc28 and Cdc5 on Nud1, Slk19, and Stu2 is important for proper spindle functions.
Park
Chong J
CJ
Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
Park
Jung-Eun
JE
Karpova
Tatiana S
TS
Soung
Nak-Kyun
NK
Yu
Li-Rong
LR
Song
Sukgil
S
Lee
Kyung H
KH
Xia
Xue
X
Kang
Eugene
E
Dabanoglu
Ilknur
I
Oh
Doo-Yi
DY
Zhang
James Y
JY
Kang
Young Hwi
YH
Wincovitch
Stephen
S
Huffaker
Tim C
TC
Veenstra
Timothy D
TD
McNally
James G
JG
Lee
Kyung S
KS
D017489:CDC28 Protein Kinase, S cerevisiae / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D008941:Spindle Apparatus; D012359:tRNA Methyltransferases
D018797:Cell Cycle Proteins; D008869:Microtubule-Associated Proteins; C109414:STU2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C437104:Slk19 protein, S cerevisiae; D012359:tRNA Methyltransferases; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; C081927:CDC5 protein, S cerevisiae; D017489:CDC28 Protein Kinase, S cerevisiae; D003851:Deoxyribonucleases; C077501:TRM2 protein, S cerevisiae