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Vol. 16, Issue 8, 3810-3820, August 2005

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Mutant Kinesin-2 Motor Subunits Increase Chromosome Loss

Mark S. Miller ^* , Jessica M. Esparza  , Andrew M. Lippa  , Fordyce G. Lux, III ^|| ¶, Douglas G. Cole ^*, and Susan K. Dutcher  ^||

^* Department of Microbiology, Molecular Biology, and Biochemistry, University of Idaho, Moscow, ID 83844-3052; Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110; and ^|| Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309-0349

Submitted May 9, 2005; Revised May 26, 2005; Accepted May 31, 2005
Monitoring Editor: Ted Salmon

The Chlamydomonas anterograde intraflagellar transport motor, kinesin-2, is isolated as a heterotrimeric complex containing two motor subunits and a nonmotor subunit known as kinesin-associated polypeptide or KAP. One of the two motor subunits is encoded by the FLA10 gene. The sequence of the second motor subunit was obtained by mass spectrometry and sequencing. It shows 46.9% identity with the Fla10 motor subunit and the gene maps to linkage group XII/XIII near RPL9. The temperature-sensitive flagellar assembly mutants fla1 and fla8 are linked to this kinesin-2 motor subunit. In each strain, a unique single point mutation gives rise to a unique single amino acid substitution within the motor domain. The fla8 strain is named fla8-1 and the fla1 strain is named fla8-2. The fla8 and fla10 alleles show a chromosome loss phenotype. To analyze this chromosome loss phenotype, intragenic revertants of fla8-1, fla8-2, and fla10-14 were generated. The analysis of the mutants and the revertants demonstrates the importance of a pocket in the amino terminus of these motor subunits for both motor activity and for a novel, dominant effect on the fidelity of chromosome segregation.

These authors made equal contributions to this work.

Present address: University of Pennsylvania Medical School, Philadelphia, PA 19104

^¶ Present address: Lander University, Greenwood, SC 29649-2099.

Address correspondence to: Susan K. Dutcher (dutcher{at}genetics.wustl.edu).

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