4.8 Article

Protein Architecture of the Human Kinetochore Microtubule Attachment Site

期刊

CELL
卷 137, 期 4, 页码 672-684

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CELL PRESS
DOI: 10.1016/j.cell.2009.03.035

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资金

  1. NIH [GM24364, GM60678, GM06627, GM86877, GM44762, GM074215]
  2. Burroughs-Wellcome Fund
  3. Leukemia and Lymphoma Society [CA06927]
  4. PA Tobacco Fund

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Chromosome segregation requires assembly of kinetochores on centromeric chromatin to mediate interactions with spindle microtubules and control cell-cycle progression. To elucidate the protein architecture of human kinetochores, we developed a two-color fluorescence light microscopy method that measures average label separation, Delta, at < 5 nm accuracy. Delta analysis of 16 proteins representing core structural complexes spanning the centromeric chromatin-microtubule interface, when correlated with mechanical states of spindle-attached kinetochores, provided a nanometer-scale map of protein position and mechanical properties of protein linkages. Treatment with taxol, which suppresses microtubule dynamics and activates the spindle checkpoint, revealed a specific switch in kinetochore architecture. Cumulatively, Delta analysis revealed that compliant linkages are restricted to the proximity of chromatin, suggested a model for how the KMN (KNL1/Mis12 complex/Ndc80 complex) network provides microtubule attachment and generates pulling forces from depolymerization, and identified an intrakinetochore molecular switch that may function in controlling checkpoint activity.

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