Gamma-tubulin and the C-terminal motor domain kinesin-like protein, KLPA, function in the establishment of spindle bipolarity in Aspergillus nidulans.
Previous research has found that a gamma-tubulin mutation in Schizosaccharomyces pombe is synthetically lethal with a deletion of the C-terminal motor domain kinesin-like protein gene pkl1, but the lethality of the double mutant prevents a phenotypic analysis of the synthetic interaction. We have investigated interactions between klpA1, a deletion of an Aspergillus nidulans homolog of pkl1, and mutations in the mipA, gamma-tubulin gene. We find that klpA1 dramatically increases the cold sensitivity and slightly reduces the growth rate at all temperatures, of three mipA alleles. In synchronized cells we find that klpA1 causes a substantial but transient inhibition of the establishment of spindle bipolarity. At a restrictive temperature, mipAD123 causes a slight, transient inhibition of spindle bipolarity and a more significant inhibition of anaphase A. In the mipAD123/klpA1 strain, formation of bipolar spindles is more strongly inhibited than in the klpA1 single mutant and many spindles apparently never become bipolar. These results indicate, surprisingly, that gamma-tubulin and the klpA kinesin have overlapping roles in the establishment of spindle bipolarity. We propose a model to account for these data.
Prigozhina
N L
NL
Department of Molecular Genetics, Ohio State University, Columbus, OH 43210, USA.
Walker
R A
RA
Oakley
C E
CE
Oakley
B R
BR
D000483:Alleles; D000705:Anaphase / Q000502:physiology; D001233:Aspergillus nidulans / Q000166:cytology* / Q000235:genetics*; D002467:Cell Nucleus; D003080:Cold Temperature; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D017353:Gene Deletion; D016547:Kinesins / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism; D008938:Mitosis / Q000502:physiology*; D009154:Mutation / Q000235:genetics / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D029702:Schizosaccharomyces pombe Proteins; D008941:Spindle Apparatus / Q000502:physiology*; D014404:Tubulin / Q000235:genetics* / Q000378:metabolism*
D005656:Fungal Proteins; C444432:KLPA protein, Aspergillus nidulans; D008869:Microtubule-Associated Proteins; C104160:PKL1 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D014404:Tubulin; C070255:mipA protein, Emericella nidulans; D016547:Kinesins