Dynamic Localization of CLIP-170 to Microtubule Plus Ends Is Coupled to Microtubule Assembly

doi:10.1083/jcb.144.1.99

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J. Cell Biol., Volume 144, Number 1, January 11, 1999 99-112

Dynamic Localization of CLIP-170 to Microtubule Plus Ends Is Coupled to Microtubule Assembly

Georgios S. Diamantopoulos,^* Franck Perez,^* Holly V. Goodson,^* Gérard Batelier, Ronald Melki, Thomas E. Kreis,^* and Janet E. Rickard^*

^* Department of Cell Biology, Sciences III, University of Geneva, CH-1211 Geneva 4, Switzerland; and Laboratoire d'Enzymologie et Biochimie Structurales, Centre National de la Recherche Scientifique, 91198 Gif-sur-Yvette, France

CLIP-170 is a cytoplasmic linker protein that localizes to plus ends of microtubules in vivo. In this study, we have characterizedthe microtubule-binding properties of CLIP-170, to understandthe mechanism of its plus end targeting. We show that the NH₂-terminalmicrotubule-interacting domain of CLIP-170 alone localizes tomicrotubule plus ends when transfected into cells. Associationof CLIP-170 with newly-formed microtubules was observed in cellsmicroinjected with biotinylated tubulin, used as a tracer forgrowing microtubules. Using in vitro assays, association of CLIP-170with recently polymerized tubulin is also seen. Cross-linkingand sedimentation velocity experiments suggest association ofCLIP-170 with nonpolymerized tubulin. We conclude from these experimentsthat the microtubule end targeting of CLIP-170 is closely linkedto tubulinpolymerization.

Key words: CLIP-170; MAPs; microtubules; polymerization; targeting

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