Complementary DNAs of two kinesin-related genes, katB and katC, were isolated from Arabidopsis thaliana and sequenced. The carboxyl-terminal regions of the polypeptides encoded by these genes, especially the presumptive ATP-binding and microtubule-binding domains, share significant sequence homology with the mechanochemical motor domain of the kinesin heavy chain. The predicted secondary structures of KatB and KatC proteins include a large globular domain in the carboxyl-terminal region and a small globular domain in the amino-terminal region that are separated by a long alpha-helical coiled-coil with heptad repeats. A truncated KatC polypeptide (KatC(207-754)), which includes the carboxyl-terminal region of KatC, was expressed in Escherichia coli and was shown to possess microtubule-stimulated ATPase activity and to bind to microtubules in an ATP-sensitive manner, both of which are characteristics of kinesin and kinesin-like proteins.