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.

[1]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[2]  E. Raff,et al.  Evidence that the head of kinesin is sufficient for force generation and motility in vitro. , 1990, Science.

[3]  N. Morris,et al.  Mutation of a gene that encodes a kinesin-like protein blocks nuclear division in A. nidulans , 1990, Cell.

[4]  B. Brinkley,et al.  Mitosis : molecules and mechanisms , 1989 .

[5]  L. Goldstein,et al.  The kinesin superfamily: tails of functional redundancy. , 1991, Trends in cell biology.

[6]  S. Inoué.  Cell division and the mitotic spindle , 1981, The Journal of cell biology.

[7]  M. Rout,et al.  Components of the yeast spindle and spindle pole body , 1990, The Journal of cell biology.

[8]  H. McDonald,et al.  Identification and characterization of a gene encoding a kinesin-like protein in Drosophila , 1990, Cell.

[9]  P. Meluh,et al.  Multiple kinesin-related proteins in yeast mitosis. , 1991, Cold Spring Harbor symposia on quantitative biology.

[10]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[11]  R. Nicklas,et al.  The forces that move chromosomes in mitosis. , 1988, Annual review of biophysics and biophysical chemistry.

[12]  P. Meluh,et al.  KAR3, a kinesin-related gene required for yeast nuclear fusion , 1990, Cell.

[13]  R. Stewart,et al.  Identification and partial characterization of six members of the kinesin superfamily in Drosophila. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. A. Laymon,et al.  A three-domain structure of kinesin heavy chain revealed by DNA sequence and microtubule binding analyses , 1989, Cell.

[15]  C. Yanisch-Perron,et al.  Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. , 1985, Gene.

[16]  S. Henikoff,et al.  Mediation of meiotic and early mitotic chromosome segregation in Drosophila by a protein related to kinesin , 1990, Nature.

[17]  P. Meluh,et al.  Suppression of the bimC4 mitotic spindle defect by deletion of klpA, a gene encoding a KAR3-related kinesin-like protein in Aspergillus nidulans , 1993, The Journal of cell biology.

[18]  Timothy J. Mitchison,et al.  Mitotic spindle organization by a plus-end-directed microtubule motor , 1992, Nature.

[19]  P. Meluh,et al.  Kinesin-related proteins required for assembly of the mitotic spindle , 1992, The Journal of cell biology.

[20]  M. Yanagida,et al.  Novel potential mitotic motor protein encoded by the fission yeast cut7+ gene , 1990, Nature.

[21]  M. Hoyt,et al.  Kinesin-related proteins required for structural integrity of the mitotic spindle , 1992, Cell.

[22]  R. Rothstein.  Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. , 1991, Methods in enzymology.

[23]  K. Loo,et al.  Two Saccharomyces cerevisiae kinesin-related gene products required for mitotic spindle assembly , 1992, The Journal of cell biology.

[24]  J. Paris,et al.  Cloning by differential screening of a Xenopus cDNA that encodes a kinesin-related protein , 1991, Molecular and cellular biology.

[25]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.