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The Mitotic Spindle

Volume 1413 of the series Methods in Molecular Biology pp 147-168

Date:

Measuring Kinetochore–Microtubule Attachment Stability in Cultured Cells

  • Keith F. DeLucaAffiliated withDepartment of Biochemistry and Molecular Biology, Colorado State University
  • , Jacob A. HermanAffiliated withDepartment of Biochemistry and Molecular Biology, Colorado State University
  • , Jennifer G. DeLucaAffiliated withDepartment of Biochemistry and Molecular Biology, Colorado State University Email author 

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Abstract

Duplicated sister chromatids connect to the mitotic spindle through kinetochores, large proteinaceous structures built at sites of centromeric heterochromatin. Kinetochores are responsible for harnessing the forces generated by microtubule polymerization and depolymerization to power chromosome movements. The fidelity of chromosome segregation relies on proper kinetochore function, as precise regulation of the attachment between kinetochores and microtubules is essential to prevent mitotic errors, which are linked to the initiation and progression of cancer and the formation of birth defects (Godek et al., Nat Rev Mol Cell Biol 16(1):57–64, 2014; Ricke and van Deursen, Semin Cell Dev Biol 22(6):559–565, 2011; Holland and Cleveland, EMBO Rep 13(6):501–514, 2012). Here we describe assays to quantitatively measure kinetochore–microtubule attachment stability in cultured cells.

Key words

Kinetochore Microtubule Spindle Mitosis Chromosome