TY - JOUR
T1 - Ndc80 internal loop interacts with Dis1/TOG to ensure proper kinetochore-spindle attachment in fission yeast
AU - Hsu, Kuo Shun
AU - Toda, Takashi
N1 - Funding Information:
We thank Fred Chang, Usula Fleig, Silke Hauf, Yasushi Hiraoka, Jonathan Millar, Kayoko Tanaka and Yoshinori Watanabe, Ayumu Yamamoto, and Mitsuhiro Yanagida for the gift of reagents used in this study and Martin Singleton's laboratory (Macromolecular Structure and Function Laboratory, Cancer Research UK, London Research Institute) for help. We are grateful to Nicola O'Reilly and the Peptide Synthesis unit for preparations of the peptide arrays. We thank Tomoyuki Tanaka for informing us of his results on budding yeast Ndc80 and Dam1 prior to publication. We are grateful to Martin Singleton for critical reading of the manuscript and stimulating discussions. This work was supported by Cancer Research UK (T.T.).
PY - 2011/2/8
Y1 - 2011/2/8
N2 - The Ndc80 complex, a conserved outer kinetochore complex, comprising four components (Ndc80/Hec1, Nuf2, Spc24, and Spc25), constitutes one of the core microtubule-binding sites within the kinetochore [1-3]. Despite this knowledge, molecular mechanisms by which this complex contributes to establishment of correct bipolar attachment of the kinetochore to the spindle microtubule remain largely elusive [1, 2, 4, 5]. Here we show that the conserved internal loop [6, 7] of fission yeast Ndc80 directly binds the Dis1/TOG microtubule-associated protein [8-10], thereby coupling spindle microtubule dynamics with kinetochore capture. Ndc80 loop mutant proteins fail to recruit Dis1 to kinetochores, imposing unstable attachment and frequent spindle collapse. In these mutants, mitotic progression is halted attributable to spindle assembly checkpoint activation, and chromosomes remain in the vicinity of the spindle poles without congression. dis1 deletion precisely phenocopies the loop mutants. Intriguingly, forced targeting of Dis1 to the Ndc80 complex rescues loop mutant's defects. We propose that Ndc80 comprises two microtubule-interacting interfaces: the N-terminal region directly binds the microtubule lattice, while the internal loop interacts with the plus end of microtubules via Dis1/TOG. Therefore, our results provide a crucial insight into how the Ndc80 complex establishes stable bipolar attachment to the spindle microtubule.
AB - The Ndc80 complex, a conserved outer kinetochore complex, comprising four components (Ndc80/Hec1, Nuf2, Spc24, and Spc25), constitutes one of the core microtubule-binding sites within the kinetochore [1-3]. Despite this knowledge, molecular mechanisms by which this complex contributes to establishment of correct bipolar attachment of the kinetochore to the spindle microtubule remain largely elusive [1, 2, 4, 5]. Here we show that the conserved internal loop [6, 7] of fission yeast Ndc80 directly binds the Dis1/TOG microtubule-associated protein [8-10], thereby coupling spindle microtubule dynamics with kinetochore capture. Ndc80 loop mutant proteins fail to recruit Dis1 to kinetochores, imposing unstable attachment and frequent spindle collapse. In these mutants, mitotic progression is halted attributable to spindle assembly checkpoint activation, and chromosomes remain in the vicinity of the spindle poles without congression. dis1 deletion precisely phenocopies the loop mutants. Intriguingly, forced targeting of Dis1 to the Ndc80 complex rescues loop mutant's defects. We propose that Ndc80 comprises two microtubule-interacting interfaces: the N-terminal region directly binds the microtubule lattice, while the internal loop interacts with the plus end of microtubules via Dis1/TOG. Therefore, our results provide a crucial insight into how the Ndc80 complex establishes stable bipolar attachment to the spindle microtubule.
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U2 - 10.1016/j.cub.2010.12.048
DO - 10.1016/j.cub.2010.12.048
M3 - Article
C2 - 21256022
AN - SCOPUS:79551705112
SN - 0960-9822
VL - 21
SP - 214
EP - 220
JO - Current Biology
JF - Current Biology
IS - 3
ER -