The human kinetochore Ska1 complex facilitates microtubule depolymerization-coupled motility.
Mitotic chromosome segregation requires that kinetochores attach to microtubule polymers and harness microtubule dynamics to drive chromosome movement. In budding yeast, the Dam1 complex couples kinetochores with microtubule depolymerization. However, a metazoan homolog of the Dam1 complex has not been identified. To identify proteins that play a corresponding role at the vertebrate kinetochore-microtubule interface, we isolated a three subunit human Ska1 complex, including the previously uncharacterized protein Rama1 that localizes to the outer kinetochore and spindle microtubules. Depletion of Ska1 complex subunits severely compromises proper chromosome segregation. Reconstituted Ska1 complex possesses two separable biochemical activities: direct microtubule binding through the Ska1 subunit, and microtubule-stimulated oligomerization imparted by the Rama1 subunit. The full Ska1 complex forms assemblies on microtubules that can facilitate the processive movement of microspheres along depolymerizing microtubules. In total, these results demonstrate a critical role for the Ska1 complex in interacting with dynamic microtubules at the outer kinetochore.
Welburn
Julie P I
JP
Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, Suite 401, Nine Cambridge Center, Cambridge, MA 02142, USA.
Grishchuk
Ekaterina L
EL
Backer
Chelsea B
CB
Wilson-Kubalek
Elizabeth M
EM
Yates
John R
JR
Cheeseman
Iain M
IM
D001483:Base Sequence; D002460:Cell Line; D002868:Chromosomal Proteins, Non-Histone / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D020090:Chromosome Segregation / Q000502:physiology; D005656:Fungal Proteins / Q000737:chemistry / Q000502:physiology; D006801:Humans; D018386:Kinetochores / Q000502:physiology*; D046529:Microscopy, Electron, Transmission; D008863:Microspheres; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000502:physiology; D008870:Microtubules / Q000502:physiology* / Q000648:ultrastructure; D008938:Mitosis / Q000502:physiology; D008954:Models, Biological; D009068:Movement / Q000502:physiology; D046912:Multiprotein Complexes; D011485:Protein Binding; D021122:Protein Subunits; D034741:RNA, Small Interfering / Q000235:genetics; D013045:Species Specificity
D002868:Chromosomal Proteins, Non-Histone; D005656:Fungal Proteins; D008869:Microtubule-Associated Proteins; D046912:Multiprotein Complexes; D021122:Protein Subunits; D034741:RNA, Small Interfering; C516427:SKA1 protein, human