Dynamic localization of CLIP-170 to microtubule plus ends is coupled to microtubule assembly

Abstract

CLIP-170 is a cytoplasmic linker protein that localizes to plus ends of microtubules in vivo. In this study, we have characterized the microtubule- binding properties of CLIP-170, to understand the mechanism of its plus end targeting. We show that the NH2-terminal microtubule-interacting domain of CLIP-170 alone localizes to microtubule plus ends when transfected into cells. Association of CLIP-170 with newly-formed microtubules was observed in cells microinjected with biotinylated tubulin, used as a tracer for growing micro- tubules. Using in vitro assays, association of CLIP-170 with recently polymerized tubulin is also seen. Crosslinking and sedimentation velocity experiments suggest association of CLIP-170 with nonpolymerized tubulin. We conclude from these experiments that the microtubule end targeting of CLIP- 170 is closely linked to tubulin polymerization.