Worm Breeder's Gazette 14(4): 24 (October 1, 1996)
Lab. of Molecular Biology Toyohashi university of Technology Toyohashi 441, Japan.
We have characterized more than a dozen members of kinesin superfamily of the motor proteins, based on the data and clones obtained from sequencing project (Y. Kohara and Genomic consortium). One of these, klp-3 has an unusual of the motor domain (which is typically found in the N-terminus region), located in the C-terminal region. Primary structure analysis shows that its length is 598 aa, it contains consensus ATP and MT binding sites at the C-terminal region. Secondary structure analysis indicates that this protein contains three distinct regions, a C-terminal globular motor domain, a coiled-coil rod and the tail which is not globular like typical kinesins. In the C. elegans so far there is only one C-terminal motor containing kinesin. To see the expression, a klp-3::lacZ construct was made. HindIII/BglII 3.5 kb fragment, containing about 1kb of the klp-3 promotor region, was put into the HindIII/BamHI site of the vector pPD16.51. The klp-3::lacZ construct was coinjected with the rol-6 marker DNA into the wild-type animals, the germ line transformants were histochemically stained for the beta-galactosidase activity. The staining is seen in pharynx, intestinal muscle and in the rectal sphincter. In the pharynx it apparently expresses in the mc1 and mc2 marginal cells (Leon Avery kindly helped us in cell identification). Previously we have reported in the west coast meeting abstract that the klp-3 (located on cosmid T09A5) can rescue the him-14 mutant phenotype partially, i. e., the number of males in the transformed progeny are reduced significantly. Also, if the klp-3 gene is injected into the wild-type animals, resulting transformants show sick worms, and about 5-10% males in the progeny. Our two factor cross data places him-14 very close to another kinesin gene unc-104, and thus it can not be in the same region as T09A5. Thus, although Klp-3 is not encoded by the him-14 gene, our results suggest that klp-3 and him-14 genes have some strong genetic interactions. Ken Kemphues and Ann Villeneuve have cloned the him-14 gene by rescueing with the cosmid ZK1127. We are interested to unravel the molecular and genetic basis of these interactions. We would like to thank the C. elegans genome sequencing consortium, Leon Avery, Y. Kohara, Ken Kemphues, Ann Villeneuve, A. Fire, M. Y. Ali, and the members of siddiqui Lab for their help.