Intrakinetochore localization and essential functional domains of Drosophila Spc105

摘要

The kinetochore is assembled during mitotic and meiotic divisions within the centromeric region of chromosomes. It is composed of more than eighty different proteins. Spc105 (also designated as Spc7, KNL-1 or Blinkin in different eukaryotes) is a comparatively large kinetochore protein, which can bind to the Mis12/MIND and Ndc80 complexes and to the spindle assembly checkpoint components Bub1 and BubR1. Our genetic characterization of Drosophila Spc105 shows that a truncated version lacking the rapidly evolving, repetitive central third still provides all essential functions. Moreover, in comparison with Cenp-C that has previously been observed to extend from the inner to the outer kinetochore region, full-length Spc105 is positioned further out and is not similarly extended along the spindle axis. Thus, our results indicate that Spc105 forms neither an extended link connecting inner Cenp-A chromatin with outer kinetochore regions nor a scaffold constraining kinetochore subcomplexes and spindle assembly checkpoint components together into a geometrically rigid supercomplex. Spc105 seems to provide a platform within the outer kinetochore allowing independent assembly of various kinetochore components.