BubR1 phosphorylates CENP-E as a switch enabling the transition from lateral association to end-on capture of spindle microtubules

Cell Res. 2019 Jul;29(7):562-578. doi: 10.1038/s41422-019-0178-z.

Yuejia Huang ^# ¹, Lin Lin ^# ² ³, Xing Liu ^# ¹ ⁴, Sheng Ye ^# ², Phil Y Yao ⁴, Wenwen Wang ¹ ⁴, Fengrui Yang ¹ ⁴, Xinjiao Gao ¹, Junying Li ¹, Yin Zhang ¹ ⁵, Jiancun Zhang ¹, Zhihong Yang ¹, Xu Liu ¹ ⁴, Zhenye Yang ¹, Jianye Zang ¹, Maikun Teng ¹, Zhiyong Wang ¹, Ke Ruan ¹, Xia Ding ⁴ ⁵, Lin Li ¹ ³, Don W Cleveland ⁶, Rongguang Zhang ⁷ ⁸, Xuebiao Yao ⁹

Affiliations

1 Anhui Key Laboratory for Chemical Biology & MOE Key Laboratory for Cellular Dynamics, CAS Center for Excellence in Molecular Cell Science, School of Life Sciences, Hefei National Laboratory for Physical Sciences at the Mciroscale, University of Science & Technology of China, Hefei, Anhui, 230026, China.
2 National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
3 National Center for Protein Science Shanghai, Institute for Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China.
4 Department of Physiology, Morehouse School of Medicine, Atlanta, GA, 30310, USA.
5 School of Basic Medical Sciences, Beijing University of Chinese Medicine, Beijing, 100029, China.
6 Ludwig Institute for Cancer Research, University of California, La Jolla, CA, 92093, USA.
7 National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China. [email protected].
8 National Center for Protein Science Shanghai, Institute for Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200031, China. [email protected].
9 Anhui Key Laboratory for Chemical Biology & MOE Key Laboratory for Cellular Dynamics, CAS Center for Excellence in Molecular Cell Science, School of Life Sciences, Hefei National Laboratory for Physical Sciences at the Mciroscale, University of Science & Technology of China, Hefei, Anhui, 230026, China. [email protected].
# Contributed equally.

PMID: 31201382 DOI: 10.1038/s41422-019-0178-z

Abstract

Error-free Mitosis depends on accurate chromosome attachment to spindle microtubules, powered congression of those chromosomes, their segregation in anaphase, and assembly of a spindle midzone at mitotic exit. The centromere-associated Kinesin motor CENP-E, whose binding partner is BubR1, has been implicated in congression of misaligned chromosomes and the transition from lateral kinetochore-microtubule association to end-on capture. Although previously proposed to be a pseudokinase, here we report the structure of the kinase domain of Drosophila melanogaster BubR1, revealing its folding into a conformation predicted to be catalytically active. BubR1 is shown to be a bona fide kinase whose phosphorylation of CENP-E switches it from a laterally attached microtubule motor to a plus-end microtubule tip tracker. Computational modeling is used to identify bubristatin as a selective BubR1 kinase antagonist that targets the αN1 helix of N-terminal extension and αC helix of the BubR1 kinase domain. Inhibition of CENP-E phosphorylation is shown to prevent proper microtubule capture at kinetochores and, surprisingly, proper assembly of the central spindle at mitotic exit. Thus, BubR1-mediated CENP-E phosphorylation produces a temporal switch that enables transition from lateral to end-on microtubule capture and organization of microtubules into stable midzone arrays.

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