title: The Dam1 ring binds to the E-hook of tubulin and diffuses along the microtubule.
abstract: There has been much effort in recent years aimed at understanding the molecular mechanism by which the Dam1 kinetochore complex is able to couple microtubule depolymerization to poleward movement. Both a biased diffusion and a forced walk model have been proposed, and several key functional aspects of Dam1-microtubule binding are disputed. Here, we investigate the elements involved in tubulin-Dam1 complex interactions and directly visualize Dam1 rings on microtubules in order to infer their dynamic behavior on the microtubule lattice and its likely relevance at the kinetochore. We find that the Dam1 complex has a preference for native tubulin over tubulin that is lacking its acidic C-terminal tail. Statistical mechanical analysis of images of Dam1 rings on microtubules, applied to both the distance between rings and the tilt angle of the rings with respect to the microtubule axis, supports a diffusive ring model. We also present a cryo-EM reconstruction of the Dam1 ring, likely the relevant assembly form of the complex for energy coupling during microtubule depolymerization in budding yeast. The present studies constitute a significant step forward by linking structural and biochemical observations toward a comprehensive understanding of the Dam1 complex.
authors.affiliation: Biophysics Graduate Group, University of California, Berkeley, Berkeley, CA 94720, USA.
authors.forename: Vincent H
authors.initials: VH
authors.lastname: Ramey
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Wang
authors.forename: Yuko
authors.initials: Y
authors.lastname: Nakajima
authors.forename: Amanda
authors.initials: A
authors.lastname: Wong
authors.forename: Jian
authors.initials: J
authors.lastname: Liu
authors.forename: David
authors.initials: D
authors.lastname: Drubin
authors.forename: Georjana
authors.initials: G
authors.lastname: Barnes
authors.forename: Eva
authors.initials: E
authors.lastname: Nogales
mesh_terms: D020090:Chromosome Segregation / Q000235:genetics; D004058:Diffusion; D018386:Kinetochores / Q000737:chemistry; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000235:genetics; D008870:Microtubules / Q000737:chemistry* / Q000235:genetics; D008956:Models, Chemical; D009363:Neoplasm Proteins / Q000737:chemistry* / Q000235:genetics*; D011485:Protein Binding / Q000235:genetics; D004718:Saccharomycetales / Q000235:genetics / Q000378:metabolism; D014404:Tubulin / Q000737:chemistry* / Q000235:genetics
chemical_list: C120624:BCAS2 protein, human; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D014404:Tubulin