title: Measuring Kinetochore-Microtubule Attachment Stability in Cultured Cells.
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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Colorado State University, MRB 237, Fort Collins, CO, 80523, USA.
authors.forename: Keith F
authors.initials: KF
authors.lastname: DeLuca
authors.affiliation: Department of Biochemistry and Molecular Biology, Colorado State University, MRB 237, Fort Collins, CO, 80523, USA.
authors.forename: Jacob A
authors.initials: JA
authors.lastname: Herman
authors.affiliation: Department of Biochemistry and Molecular Biology, Colorado State University, MRB 237, Fort Collins, CO, 80523, USA. jdeluca@colostate.edu.
authors.forename: Jennifer G
authors.initials: JG
authors.lastname: DeLuca
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D020868:Gene Silencing; D006801:Humans; D018386:Kinetochores / Q000737:chemistry / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D055503:Protein Multimerization; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D008941:Spindle Apparatus / Q000737:chemistry / Q000378:metabolism; D013696:Temperature
chemical_list: D009687:Nuclear Proteins; D034741:RNA, Small Interfering
keywords: Chromosome; Kinetochore; Microtubule; Mitosis; Spindle