Mitotic phosphorylation by Nek6 and Nek7 reduces microtubule affinity of EML4 to alter spindle dynamics and promote chromosome congression

Fry, A. M.; Adib, R.; Montgomery, J. M.; Atherton, J.; O'Regan, L.; Richards, M.; Straatman, K. R.; Roth, D.; Straube, A.; Bayliss, R.; Moores, C. A.

doi:10.1101/466979

bioRxiv · 10.1101/466979

Mitotic phosphorylation by Nek6 and Nek7 reduces microtubule affinity of EML4 to alter spindle dynamics and promote chromosome congression

Fry, A. M.·Adib, R.·Montgomery, J. M.·Atherton, J.·O'Regan, L.·Richards, M.·Straatman, K. R.·Roth, D.·Straube, A.·Bayliss, R.·Moores, C. A.

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Abstract

EML4 is a microtubule-associated protein that promotes microtubule stability. We show here that EML4 is distributed as punctate foci along the microtubule lattice in interphase but exhibits reduced association with spindle microtubules in mitosis. Microtubule sedimentation and cryo-electron microscopy and 3D reconstruction reveal that EML4 binds via its basic N-terminal domain to the acidic C-terminal tails of - and {beta}-tubulin on the microtubule surface. The mitotic kinases Nek6 and Nek7 can phosphorylate EML4 N-terminal domain at S144 and S146 in vitro, and depletion of these kinases leads to increased EML4 binding to microtubules in mitosis. An S144A-S146A double mutant not only binds inappropriately to mitotic microtubules but also interferes with chromosome congression. Meanwhile, constitutive activation of Nek6 or Nek7 reduces EML4 association with interphase microtubules. Together, these data support a model in which Nek6 and Nek7-dependent phosphorylation promotes dissociation of EML4 from microtubules in mitosis thereby altering microtubule dynamics to enable chromosome congression.

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Fry, A. M., Adib, R., Montgomery, J. M., Atherton, J., O'Regan, L., Richards, M., Straatman, K. R., Roth, D., Straube, A., Bayliss, R., Moores, C. A.. 2018-11-09. Mitotic phosphorylation by Nek6 and Nek7 reduces microtubule affinity of EML4 to alter spindle dynamics and promote chromosome congression. https://doi.org/10.1101/466979

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