title: Merotelic kinetochore orientation is a major mechanism of aneuploidy in mitotic mammalian tissue cells.
abstract: In mitotic cells, an error in chromosome segregation occurs when a chromosome is left near the spindle equator after anaphase onset (lagging chromosome). In PtK1 cells, we found 1.16% of untreated anaphase cells exhibiting lagging chromosomes at the spindle equator, and this percentage was enhanced to 17.55% after a mitotic block with 2 microM nocodazole. A lagging chromosome seen during anaphase in control or nocodazole-treated cells was found by confocal immunofluorescence microscopy to be a single chromatid with its kinetochore attached to kinetochore microtubule bundles extending toward opposite poles. This merotelic orientation was verified by electron microscopy. The single kinetochores of lagging chromosomes in anaphase were stretched laterally (1.2--5.6-fold) in the directions of their kinetochore microtubules, indicating that they were not able to achieve anaphase poleward movement because of pulling forces toward opposite poles. They also had inactivated mitotic spindle checkpoint activities since they did not label with either Mad2 or 3F3/2 antibodies. Thus, for mammalian cultured cells, kinetochore merotelic orientation is a major mechanism of aneuploidy not detected by the mitotic spindle checkpoint. The expanded and curved crescent morphology exhibited by kinetochores during nocodazole treatment may promote the high incidence of kinetochore merotelic orientation that occurs after nocodazole washout.
authors.affiliation: Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: D
authors.initials: D
authors.lastname: Cimini
authors.forename: B
authors.initials: B
authors.lastname: Howell
authors.forename: P
authors.initials: P
authors.lastname: Maddox
authors.forename: A
authors.initials: A
authors.lastname: Khodjakov
authors.forename: F
authors.initials: F
authors.lastname: Degrassi
authors.forename: E D
authors.initials: ED
authors.lastname: Salmon
mesh_terms: D000705:Anaphase; D000782:Aneuploidy; D000818:Animals; D002135:Calcium-Binding Proteins / Q000302:isolation & purification; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D016764:Cell Polarity; D002503:Centromere / Q000502:physiology / Q000648:ultrastructure; D002842:Chromatids / Q000502:physiology; D002875:Chromosomes / Q000502:physiology; D000939:Epitopes; D005656:Fungal Proteins / Q000302:isolation & purification; D018386:Kinetochores / Q000502:physiology* / Q000648:ultrastructure; D008870:Microtubules / Q000502:physiology; D008938:Mitosis / Q000187:drug effects / Q000502:physiology*; D008957:Models, Genetic; D008961:Models, Structural; D009068:Movement; D015739:Nocodazole / Q000494:pharmacology; D009687:Nuclear Proteins; D008941:Spindle Apparatus / Q000502:physiology; D013692:Telophase
chemical_list: D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D000939:Epitopes; D005656:Fungal Proteins; D009687:Nuclear Proteins; D015739:Nocodazole