The XMAP215-family protein DdCP224 is required for cortical interactions of microtubules.
BACKGROUND
Interactions of peripheral microtubule tips with the cell cortex are of crucial importance for nuclear migration, spindle orientation, centrosome positioning and directional cell movement. Microtubule plus end binding proteins are thought to mediate interactions of microtubule tips with cortical actin and membrane proteins in a dynein-dependent manner. XMAP215-family proteins are main regulators of microtubule plus end dynamics but so far they have not been implicated in the interactions of microtubule tips with the cell cortex.


RESULTS
Here we show that overexpression of an N-terminal fragment of DdCP224, the Dictyostelium XMAP215 homologue, caused a collapse of the radial microtubule cytoskeleton, whereby microtubules lost contact with the cell cortex and were dragged behind like a comet tail of an unusually motile centrosome. This phenotype was indistinguishable from mutants overexpressing fragments of the dynein heavy chain or intermediate chain. Moreover, it was accompanied by dispersal of the Golgi apparatus and reduced cortical localization of the dynein heavy chain indicating a disrupted dynein/dynactin interaction. The interference of DdCP224 with cortical dynein function is strongly supported by the observations that DdCP224 and its N-terminal fragment colocalize with dynein and coimmunoprecipitate with dynein and dynactin.


CONCLUSIONS
Our data show that XMAP215-like proteins are required for the interaction of microtubule plus ends with the cell cortex in interphase cells and strongly suggest that this function is mediated by dynein.
Hestermann
Andrea
A
A,-Butenandt-Institut/Zellbiologie, Ludwig-Maximilians-Universität München, Schillerstr, 42, D-80336 München, Germany. andrea.hestermann@lrz.uni-muenchen.de
Gräf
Ralph
R
D000818:Animals; D004023:Dictyostelium / Q000648:ultrastructure*; D008869:Microtubule-Associated Proteins / Q000145:classification / Q000502:physiology*; D008870:Microtubules / Q000648:ultrastructure*; D015800:Protozoan Proteins / Q000145:classification / Q000502:physiology*; D029867:Xenopus Proteins / Q000145:classification
C486738:CKAP5 protein, Xenopus; D008869:Microtubule-Associated Proteins; D015800:Protozoan Proteins; D029867:Xenopus Proteins; C411383:centrosomal protein 224-kDa, Dictyostelium