Golgi/TGN cytoplasmic linker protein belonging to the CLIP-170 family

he microtubule cytoskeleton plays a fundamental role in cell organization and membrane traffic in higher eukaryotes. It is well established that molecular motors are involved in membrane–microtubule interactions, but it has also been proposed that nonmotor microtubulebinding (MTB) proteins known as CLIPs (cytoplasmic linker proteins) have basic roles in these processes. We report here the characterization of CLIPR-59, a CLIP-170–related protein localized to the trans-most part of the Golgi apparatus. CLIPR-59 contains an acidic region followed by three ankyrin-like repeats and two CLIP-170–related MTB motifs. We show that the 60–amino acid–long carboxy-terminal domain of CLIPR-59 is necessary and sufficient to achieve T Golgi targeting, which represents the first identification of a membrane targeting domain in a CLIP-170–related protein. The MTB domain of CLIPR-59 is functional because it localizes to microtubules when expressed as a fragment in HeLa cells. However, our results suggest that this domain is normally inhibited by the presence of adjacent domains, because neither full-length CLIPR-59 nor a CLIPR-59 mutant missing its membrane-targeting region localize to microtubules. Consistent with this observation, overexpression of CLIPR-59 does not affect the microtubule network. However, CLIPR-59 overexpression strongly perturbs early/recycling endosome–TGN dynamics, implicating CLIPR-59 in the regulation of this pathway.