The Dam1 kinetochore ring complex moves processively on depolymerizing microtubule ends.
Chromosomes interact through their kinetochores with microtubule plus ends and they are segregated to the spindle poles as the kinetochore microtubules shorten during anaphase A of mitosis. The molecular natures and identities of coupling proteins that allow microtubule depolymerization to pull chromosomes to poles during anaphase have long remained elusive. In budding yeast, the ten-protein Dam1 complex is a critical microtubule-binding component of the kinetochore that oligomerizes into a 50-nm ring around a microtubule in vitro. Here we show, with the use of a real-time, two-colour fluorescence microscopy assay, that the ring complex moves processively for several micrometres at the ends of depolymerizing microtubules without detaching from the lattice. Electron microscopic analysis of 'end-on views' revealed a 16-fold symmetry of the kinetochore rings. This out-of-register arrangement with respect to the 13-fold microtubule symmetry is consistent with a sliding mechanism based on an electrostatically coupled ring-microtubule interface. The Dam1 ring complex is a molecular device that can translate the force generated by microtubule depolymerization into movement along the lattice to facilitate chromosome segregation.
Westermann
Stefan
S
Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California 94720-3202, USA.
Wang
Hong-Wei
HW
Avila-Sakar
Agustin
A
Drubin
David G
DG
Nogales
Eva
E
Barnes
Georjana
G
D018797:Cell Cycle Proteins / Q000502:physiology*; D020090:Chromosome Segregation / Q000502:physiology; D018386:Kinetochores / Q000502:physiology* / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000502:physiology*; D008870:Microtubules / Q000502:physiology* / Q000648:ultrastructure; D009068:Movement; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology*; D008941:Spindle Apparatus / Q000502:physiology* / Q000648:ultrastructure
D018797:Cell Cycle Proteins; C116003:DAM1 protein, S cerevisiae; D008869:Microtubule-Associated Proteins; D029701:Saccharomyces cerevisiae Proteins