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Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
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Multisite phosphorylation of the NDC80 complex gradually tunes its microtubule-binding affinity
Anatoly V Zaytsev1, Jeanne E Mick2, Evgeny Maslennikov3
1Physiology Department, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104.
Molecular Biology of the Cell
|March 27, 2015
Summary
Phosphorylation of the NDC80 complex gradually tunes microtubule (MT) binding, acting as a rheostat. This regulation by Hec1 tail phosphorylation impacts cell division dynamics.
Area of Science:
- Cell Biology
- Biophysics
- Molecular Biology
Background:
- Microtubule (MT) attachment to kinetochores is essential for accurate cell division.
- The precise mechanisms by which phosphorylation regulates these crucial interactions remain largely unknown.
Purpose of the Study:
- To investigate the phosphoregulation of the NDC80 complex, a key kinetochore component.
- To elucidate how phosphorylation events on the Hec1 subunit's tail control NDC80-MT binding.
Main Methods:
- Quantitative in vitro biochemical assays.
- Molecular dynamics simulations.
- Analysis of Hec1 subunit phosphorylation.
Main Results:
- Multiple phosphorylation events on the Hec1 tail are additively integrated, gradually tuning NDC80-MT binding.
- The Hec1 tail acts as a phosphorylation-controlled rheostat, modulating MT binder affinity.
- NDC80 complex cooperativity is weak and unaffected by phosphorylation.
Conclusions:
- Phosphorylation provides a mechanism for fine-tuning kinetochore-MT attachments.
- Independent binding events of individual NDC80 complexes, not oligomers, regulate kinetochore-MT dynamics and stability during cell division.
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