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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 457-466 |
| Number of pages | 10 |
| Journal | Molecular Biology of the Cell |
| Volume | 22 |
| Issue number | 4 |
| DOIs | |
| State | Published - Feb 15 2011 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Molecular Biology
- Cell Biology
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The Dam1 ring binds to the E-hook of tubulin and diffuses along the microtubule. / Ramey, Vincent H.; Wang, Hong Wei; Nakajima, Yuko; Wong, Amanda; Liu, Jian; Drubin, David; Barnes, Georjana; Nogales, Eva.
In: Molecular Biology of the Cell, Vol. 22, No. 4, 15.02.2011, p. 457-466.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - The Dam1 ring binds to the E-hook of tubulin and diffuses along the microtubule
AU - Ramey, Vincent H.
AU - Wang, Hong Wei
AU - Nakajima, Yuko
AU - Wong, Amanda
AU - Liu, Jian
AU - Drubin, David
AU - Barnes, Georjana
AU - Nogales, Eva
PY - 2011/2/15
Y1 - 2011/2/15
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=79951826786&partnerID=8YFLogxK
U2 - 10.1091/mbc.E10-10-0841
DO - 10.1091/mbc.E10-10-0841
M3 - Article
C2 - 21169562
AN - SCOPUS:79951826786
VL - 22
SP - 457
EP - 466
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
SN - 1059-1524
IS - 4
ER -