Dynamic crotonylation of EB1 by TIP60 ensures accurate spindle positioning in mitosis

Xiaoyu Song; Fengrui Yang; Xu Liu; Peng Xia; Wu Yin; Zhikai Wang; Yong Wang; Xiao Yuan; Zhen Dou; Kai Jiang; Mingming Ma; Bing Hu; Rui Zhang; Chao Xu; Zhiyong Zhang; Ke Ruan; Ruijun Tian; Lin Li; Tao Liu; Donald L Hill; Jianye Zang; Xing Liu; Jinsong Li; Jinke Cheng; Xuebiao Yao

doi:10.1038/s41589-021-00875-7

Dynamic crotonylation of EB1 by TIP60 ensures accurate spindle positioning in mitosis

Nat Chem Biol. 2021 Dec;17(12):1314-1323. doi: 10.1038/s41589-021-00875-7. Epub 2021 Oct 4.

Authors

Xiaoyu Song^#^{1

2}, Fengrui Yang^#^{1

2}, Xu Liu^#^{1

2}, Peng Xia^#¹, Wu Yin¹, Zhikai Wang^{1

2}, Yong Wang³, Xiao Yuan^{1

4}, Zhen Dou¹, Kai Jiang³, Mingming Ma¹, Bing Hu¹, Rui Zhang⁵, Chao Xu¹, Zhiyong Zhang¹, Ke Ruan¹, Ruijun Tian⁴, Lin Li⁶, Tao Liu⁷, Donald L Hill⁸, Jianye Zang⁹, Xing Liu^{10

11}, Jinsong Li¹², Jinke Cheng¹³, Xuebiao Yao^{14

15

16}

Affiliations

¹ MOE Key Laboratory for Cellular Dynamics and Hefei National Center for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei, China.
² Keck Center for Organoids Plasticity, Morehouse School of Medicine, Atlanta, GA, USA.
³ Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Wuhan University, Wuhan, China.
⁴ Department of Chemistry, Southern University of Science and Technology, Shenzhen, China.
⁵ Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO, USA.
⁶ Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China.
⁷ State Key Laboratory of Natural and Biomimetic Drugs, Peking University, Beijing, China.
⁸ Comprehensive Cancer Center, University of Alabama, Birmingham, AL, USA.
⁹ MOE Key Laboratory for Cellular Dynamics and Hefei National Center for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei, China. [email protected].
¹⁰ MOE Key Laboratory for Cellular Dynamics and Hefei National Center for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei, China. [email protected].
¹¹ Keck Center for Organoids Plasticity, Morehouse School of Medicine, Atlanta, GA, USA. [email protected].
¹² Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China. [email protected].
¹³ Department of Biochemistry, Shanghai Jiaotong University School of Medicine, Shanghai, China. [email protected].
¹⁴ MOE Key Laboratory for Cellular Dynamics and Hefei National Center for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei, China. [email protected].
¹⁵ Keck Center for Organoids Plasticity, Morehouse School of Medicine, Atlanta, GA, USA. [email protected].
¹⁶ Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China. [email protected].

^# Contributed equally.

PMID: 34608293
DOI: 10.1038/s41589-021-00875-7

Abstract

Spindle position control is essential for cell fate determination and organogenesis. Early studies indicate the essential role of the evolutionarily conserved Gαi/LGN/NuMA network in spindle positioning. However, the regulatory mechanisms that couple astral microtubules dynamics to the spindle orientation remain elusive. Here we delineated a new mitosis-specific crotonylation-regulated astral microtubule-EB1-NuMA interaction in mitosis. EB1 is a substrate of TIP60, and TIP60-dependent crotonylation of EB1 tunes accurate spindle positioning in mitosis. Mechanistically, TIP60 crotonylation of EB1 at Lys66 forms a dynamic link between accurate attachment of astral microtubules to the lateral cell cortex defined by NuMA-LGN and fine tune of spindle positioning. Real-time imaging of chromosome movements in HeLa cells expressing genetically encoded crotonylated EB1 revealed the importance of crotonylation dynamics for accurate control of spindle orientation during metaphase-anaphase transition. These findings delineate a general signaling cascade that integrates protein crotonylation with accurate spindle positioning for chromosome stability in mitosis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Cell Cycle Proteins / metabolism*
Chromosomes / ultrastructure
Escherichia coli / genetics
HeLa Cells
Humans
Kinetics
Lysine Acetyltransferase 5 / metabolism*
Microtubule-Associated Proteins / metabolism*
Microtubules / metabolism*
Mitosis
Protein Binding
Protein Conformation
Spindle Apparatus / metabolism*

Substances

Cell Cycle Proteins
MAPRE1 protein, human
Microtubule-Associated Proteins
NUMA1 protein, human
KAT5 protein, human
Lysine Acetyltransferase 5