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Updated: Jul 14, 2026

Studying Mitotic Checkpoint by Illustrating Dynamic Kinetochore Protein Behavior and Chromosome Motion in Living Drosophila Syncytial Embryos
Published on: June 14, 2012
Kinetochore dynein is required for chromosome motion and congression independent of the spindle checkpoint
Zhenye Yang1, U Serdar Tulu, Patricia Wadsworth
1Lab of Cell Growth and Division, Division of Molecular Medicine, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509, USA.
Abstract:
During mitosis, the motor molecule cytoplasmic dynein plays key direct and indirect roles in organizing microtubules (MTs) into a functional spindle. At this time, dynein is also recruited to kinetochores, but its role or roles at these organelles remain vague, partly because inhibiting dynein globally disrupts spindle assembly [1-4]. However, dynein can be selectively depleted from kinetochores by disruption of ZW10 [5], and recent studies with this approach conclude that kinetochore-associated dynein (KD) functions to silence the spindle-assembly checkpoint (SAC) [6]. Here we use dynein-antibody microinjection and the RNAi of ZW10 to explore the role of KD in chromosome behavior during mitosis in mammals. We find that depleting or inhibiting KD prevents the rapid poleward motion of attaching kinetochores but not kinetochore fiber (K fiber) formation. However, after kinetochores attach to the spindle, KD is required for stabilizing kinetochore MTs, which it probably does by generating tension on the kinetochore, and in its absence, chromosome congression is defective. Finally, depleting KD reduces the velocity of anaphase chromosome motion by approximately 40%, without affecting the rate of poleward MT flux. Thus, in addition to its role in silencing the SAC, KD is important for forming and stabilizing K fibers and in powering chromosome motion.
Insights
Kinetochore-associated dynein (KD) is crucial for chromosome alignment and movement during cell division. This motor protein stabilizes microtubules and powers chromosome motion, in addition to silencing the spindle-assembly checkpoint.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- Cytoplasmic dynein is essential for organizing microtubules into a functional spindle during mitosis.
- The specific roles of dynein at kinetochores remain unclear due to global inhibition effects.
- Kinetochore-associated dynein (KD) has been implicated in silencing the spindle-assembly checkpoint (SAC).
Purpose of the Study:
- To investigate the role of KD in chromosome behavior during mammalian mitosis.
- To explore KD's functions beyond SAC silencing.
Main Methods:
- Selective depletion of KD by disrupting ZW10.
- Dynein-antibody microinjection.
- RNA interference (RNAi) of ZW10.
Main Results:
- KD depletion prevents poleward motion of attaching kinetochores but not kinetochore fiber formation.
- KD is required for stabilizing kinetochore microtubules and generating tension after kinetochore attachment.
- Chromosome congression is defective, and anaphase chromosome motion velocity is reduced by ~40% upon KD depletion.
- Poleward microtubule flux rates remain unaffected.
Conclusions:
- KD plays vital roles in forming and stabilizing kinetochore fibers.
- KD is essential for proper chromosome congression and motion during mitosis.
- Beyond silencing the SAC, KD is critical for microtubule dynamics and chromosome movement in mammalian cells.
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