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© The Rockefeller University Press, 0021-9525/2000/3/1255/ $5.00
The Journal of Cell Biology, Volume 148, Number 6, March 20, 2000 1255-1266

Original Article

Kinesin Superfamily Protein 3 (KIF3) Motor Transports Fodrin-associating Vesicles Important for Neurite Building

Sen Takedaa, Hiroto Yamazakia, Dae-Hyun Seoga, Yoshimitsu Kanaia, Sumio Teradaa, and Nobutaka Hirokawaa
a Department of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan

Correspondence to: Nobutaka Hirokawa, Department of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. Tel:+81-3-5841-3326

Kinesin superfamily proteins (KIFs) comprise several dozen molecular motor proteins. The KIF3 heterotrimer complex is one of the most abundantly and ubiquitously expressed KIFs in mammalian cells. To unveil the functions of KIF3, microinjection of function-blocking monovalent antibodies against KIF3 into cultured superior cervical ganglion (SCG) neurons was carried out. They significantly blocked fast axonal transport and brought about inhibition of neurite extension. A yeast two-hybrid binding assay revealed the association of fodrin with the KIF3 motor through KAP3. This was further confirmed by using vesicles collected from large bundles of axons (cauda equina), from which membranous vesicles could be prepared in pure preparations. Both immunoprecipitation and immunoelectron microscopy indicated the colocalization of fodrin and KIF3 on the same vesicles, the results reinforcing the evidence that the cargo of the KIF3 motor consists of fodrin-associating vesicles. In addition, pulse-labeling study implied partial comigration of both molecules as fast flow components. Taken together, the KIF3 motor is engaged in fast axonal transport that conveys membranous components important for neurite extension.

Key Words: kinesin superfamily protein 3, fodrin, microinjection, axonal transport, yeast two-hybrid


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