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.
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20 March 2000
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March 20 2000
Kinesin Superfamily Protein 3 (Kif3) Motor Transports Fodrin-Associating Vesicles Important for Neurite Building
Sen Takeda,
Sen Takeda
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
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Hiroto Yamazaki,
Hiroto Yamazaki
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
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Dae-Hyun Seog,
Dae-Hyun Seog
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
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Yoshimitsu Kanai,
Yoshimitsu Kanai
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
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Sumio Terada,
Sumio Terada
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
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Nobutaka Hirokawa
Nobutaka Hirokawa
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
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Sen Takeda
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Hiroto Yamazaki
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Dae-Hyun Seog
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Yoshimitsu Kanai
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Sumio Terada
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Nobutaka Hirokawa
aDepartment of Cell Biology and Anatomy, University of Tokyo, Graduate School of Medicine, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
Abbreviations used in this paper: aa, amino acids; CyDy, cytoplasmic dynein; KIF, kinesin superfamily protein; MT, microtubule; SCG, superior cervical ganglion.
Received:
May 04 1999
Revision Requested:
February 07 2000
Accepted:
February 09 2000
Online ISSN: 1540-8140
Print ISSN: 0021-9525
© 2000 The Rockefeller University Press
2000
The Rockefeller University Press
J Cell Biol (2000) 148 (6): 1255–1266.
Article history
Received:
May 04 1999
Revision Requested:
February 07 2000
Accepted:
February 09 2000
Citation
Sen Takeda, Hiroto Yamazaki, Dae-Hyun Seog, Yoshimitsu Kanai, Sumio Terada, Nobutaka Hirokawa; Kinesin Superfamily Protein 3 (Kif3) Motor Transports Fodrin-Associating Vesicles Important for Neurite Building. J Cell Biol 20 March 2000; 148 (6): 1255–1266. doi: https://doi.org/10.1083/jcb.148.6.1255
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