Centrosomal microtubule plus end tracking proteins and their role in Dictyostelium cell dynamics.

Abstract

Microtubules interact with huge protein complexes not only with their minus ends but also with their peripheral plus ends. The centrosome at their minus ends nucleates and organizes the microtubule cytoskeleton. The microtubule plus end complex seems to be required for the capture of microtubule tips at cortical sites by mediating interactions of microtubule tips with cortical actin as well as with membrane proteins. This process plays a major role in nuclear migration, spindle orientation and directional cell movement. Five potential members of the microtubule plus end complex have already been identified in Dictyostelium, DdCP224, DdEB1, DdLIS1, the dynein heavy chain and dynein intermediate chain. DdCP224 and DdEB1 are the Dictyostelium representatives of the XMAP215- and EB1-family, respectively. Both are not only concentrated at microtubule tips, they are also centrosomal components. The centrosomal binding domain of DdCP224 resides within the C-terminal fifth of the protein. DdCP224 is involved in the centrosome duplication cycle and cytokinesis. DdEB1 is the first member of the EB1 protein family that is also a genuine centrosomal component. A DdEB1 null mutant revealed that DdEB1 is required for mitotic spindle formation. DdEB1 coprecipitates and colocalizes with DdCP224 suggesting that these proteins act together in their functions. One of these functions could be dynein/dynactin-dependent interaction of microtubule tips with the cell cortex that is thought to determine the positioning of the microtubule-organizing center (MTOC) and the direction of migration.