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Multivalency of NDC80 in the outer kinetochore is essential to track shortening microtubules and generate forces

Elife. 2018-04; 
Volkov VA, Huis In 't Veld PJ, Dogterom M, Musacchio A.
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PCR Cloning and Subcloning m Source or reference Identifiers Additional information GenScript GenScript Dogterom laboratory, this study peptide for C-terminal sortase labeling peptide for C-terminal sortase labeling Matlab script to trace fluorescent particles in kymographs Protein expression and purification Standard Gibson assembly or restriction-ligation dependent cloning techniques were used to gener- ate pLIB vectors with NDC80, SPC25SORT-HIS, SPC24, and SPC24SPY. Get A Quote
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摘要

Presence of multiple copies of the microtubule-binding NDC80 complex is an evolutionary conserved feature of kinetochores, points of attachment of chromosomes to spindle microtubules. This may enable multivalent attachments to microtubules, with implications that remain unexplored. Using recombinant human kinetochore components, we show that while single NDC80 complexes do not track depolymerizing microtubules, reconstituted particles containing the NDC80 receptor CENP-T bound to three or more NDC80 complexes do so effectively, as expected for a kinetochore force coupler. To study multivalency systematically, we engineered modules allowing incremental addition of NDC80 complexes. The modules' residence time on ... More

关键词

CENP-T; Hec1; Ndc80; biochemistry; chemical biology; human; kinetochore; microtubule; molecular biophysics; multivalency; structural biology
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