Cytoplasmic dynein/dynactin drives kinetochore protein transport to the spindle poles and has a role in mitotic spindle checkpoint inactivation

By B. Howell, B. McEwen, J. Canman, D. B. Hoffman, E. M. Farrar, C. Rieder, E. Salmon. Journal of Cell Biology, 2001

623 citations recorded by the source corpus.

We discovered that many proteins located in the kinetochore outer domain, but not the inner core, are depleted from kinetochores and accumulate at spindle poles when ATP production is suppressed in PtK1 cells, and that microtubule depolymerization inhibits this process. These proteins include the microtubule motors CENP-E and cytoplasmic dynein, and proteins involved with the mitotic spindle checkpoint, Mad2, Bub1R, and the 3F3/2 phosphoantigen. Depletion of these components did not disrupt kinetochore outer domain structure or alter metaphase kinetochore microtubule number. Inhibition of dynein/dynactin activity by microinjection in prometaphase with purified p50 “dynamitin” protein or concentrated 70.1 anti-dynein antibody blocked outer domain protein transport to the spindle poles, prevented Mad2 depletion from kinetochores despite normal kinetochore microtubule numbers, reduced metaphase kinetochore tension by 40%, and induced a mitotic block at metaphase. Dynein/dynactin inhibition did not block chromosome congression to the spindle equator in prometaphase, or segregation to the poles in anaphase when the spindle checkpoint was inactivated by microinjection with Mad2 antibodies. Thus, a major function of dynein/dynactin in mitosis is in a kinetochore disassembly pathway that contributes to inactivation of the spindle checkpoint.

Subfields: Cell Biology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/66e15ea2db5fe6626454d0a8461a428e60e26b68

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In Cell Biology

1. A. Musacchio
2. Stephen S. Taylor
3. E. Salmon
4. R. Vale
5. Debashish Chowdhury

In Molecular Biology

1. A. Musacchio
2. Stephen S. Taylor
3. E. Salmon
4. R. Vale
5. Julia R. Kardon

Citations in the corpus

• Regulators of the cytoplasmic dynein motor (2009, background): “Once a sister chromatid pair becomes bi-oriented, dynein transports the spindle assembly checkpoint (SAC) proteins mitotic arrest deficient-like protein 1 (MAD1), MAD2 and BUBR1, as well as RZZ and Spindly, away from the kinetochores and towards the spindle poles, thereby silencing the SAC and all”
• The spindle assembly checkpoint. (2012, background): “Stripping is mediated by the minusend directed microtubule motor dynein as inhibiting dynein function blocks removal of Mad1 and Mad2 from attached kinetochores [121].”
• The conserved KMN network constitutes the core microtubule-binding site of the kinetochore. (2006, background): “In metazoans, inhibition of the two kinetochore motors (cytoplasmic dynein and CENP-E) does not significantly alter chromosomespindle interactions for the majority of chromosomes (Howell et al., 2001; Putkey et al., 2002).”
• The spindle checkpoint: structural insights into dynamic signalling (2002, background): “If dynein is inhibited, Mad2 accumulates at kinetochores and keeps cycling with a high turnover rate, showing that distinct mechanisms regulate the interaction of Mad2 with kinetochore”
• The spindle-assembly checkpoint in space and time (2007, background): “and dynein; and dynein-associated proteins such as dynactin, cytoplasmic linker protein CLIP170 and lissencephaly-1 ( LIS1 ) (Ref”