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J. Biol. Chem., Vol. 279, Issue 18, 19201-19208, April 30, 2004

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A Direct Interaction between Cytoplasmic Dynein and Kinesin I May Coordinate Motor Activity^*

Lee A. Ligon, Mariko Tokito, Jeffrey M. Finklestein, Francesca E. Grossman, and Erika L. F. Holzbaur

From the Department of Physiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6085

Cytoplasmic dynein and kinesin I are both unidirectional intracellular motors. Dynein moves cargo toward the cell center, and kinesin moves cargo toward the cell periphery. There is growing evidence that bi-directional motility is regulated in the cell, potentially through direct interactions between oppositely oriented motors. We have identified a direct interaction between cytoplasmic dynein and kinesin I. Using the yeast two-hybrid assay and affinity chromatography, we demonstrate that the intermediate chain of dynein binds to kinesin light chains 1 and 2. The interaction is both direct and specific. Co-immunoprecipitation experiments demonstrate an interaction between endogenous proteins in rat brain cytosol. Double-label immunocytochemistry reveals a partial co-localization of vesicle-associated motor proteins. Together these observations suggest that soluble motors can interact, potentially allowing kinesin I to actively localize dynein to cellular sites of function. There is also a vesicle population with both dynein and kinesin I bound that may be capable of bi-directional motility along cellular microtubules.

Received for publication, December 9, 2003 , and in revised form, February 18, 2004.

^* This work was supported by National Institutes of Health Grant GM48661 (to E. L. F. H.) and a National Institutes of Health postdoctoral fellowship (to L. A. L.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Physiology, University of Pennsylvania School of Medicine, D400 Richards Bldg., 3700 Hamilton Walk, Philadelphia, PA 19104-6085. Tel.: 215-573-3257; Fax: 215-573-5851; E-mail: holzbaur{at}mail.med.upenn.edu.

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