Loss of function of Saccharomyces cerevisiae kinesin-related CIN8 and KIP1 is suppressed by KAR3 motor domain mutations.
The kinesin-related products of the CIN8 and KIP1 genes of Saccharomyces cerevisiae redundantly perform an essential function in mitosis. The action of either gene-product is required for an outwardly directed force that acts upon the spindle poles. We have selected mutations that suppress the temperature-sensitivity of a cin8-temperature-sensitive kip1-delta strain. The extragenic suppressors analyzed were all found to be alleles of the KAR3 gene. KAR3 encodes a distinct kinesin-related protein whose action antagonizes Cin8p/Kip1p function. All seven alleles analyzed were altered within the region of KAR3 that encodes the putative force-generating (or "motor") domain. These mutations also suppressed the inviability associated with the cin8-delta kip1-delta genotype, a property not shared by a deletion of KAR3. Other properties of the suppressing alleles revealed that they were not null for function. Six of the seven were unaffected for the essential karyogamy and meiosis properties of KAR3 and the seventh was dominant for the suppressing trait. Our findings suggest that despite an antagonistic relationship between Cin8p/Kip1p and Kar3p, aspects of their mitotic roles may be similar.
Hoyt
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MA
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.
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Totis
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Saunders
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D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D004271:DNA, Fungal; D005656:Fungal Proteins / Q000235:genetics*; D016153:Genes, Suppressor; D016547:Kinesins / Q000235:genetics*; D008540:Meiosis / Q000235:genetics; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D008938:Mitosis / Q000235:genetics; D020409:Molecular Motor Proteins; D008969:Molecular Sequence Data; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D008941:Spindle Apparatus
C075342:CIN8 protein, S cerevisiae; D004271:DNA, Fungal; D005656:Fungal Proteins; C063504:KAR3 protein, S cerevisiae; C431747:KIP1 protein, S cerevisiae; D008869:Microtubule-Associated Proteins; D020409:Molecular Motor Proteins; D029701:Saccharomyces cerevisiae Proteins; D016547:Kinesins