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Redox-dependent regulation of end-binding protein 1 activity by glutathionylation

Science China Life Sciences volume 64, pages 575–583 (2021)Cite this article

Abstract

Cytoskeletal proteins are susceptible to glutathionylation under oxidizing conditions, and oxidative damage has been implicated in several neurodegenerative diseases. End-binding protein 1 (EB1) is a master regulator of microtubule plus-end tracking proteins (+TIPs) and is critically involved in the control of microtubule dynamics and cellular processes. However, the impact of glutathionylation on EB1 functions remains unknown. Here we reveal that glutathionylation is important for controlling EB1 activity and protecting EB1 from irreversible oxidation. In vitro biochemical and cellular assays reveal that EB1 is glutathionylated. Diamide, a mild oxidizing reagent, reduces EB1 comet number and length in cells, indicating the impairment of microtubule dynamics. Three cysteine residues of EB1 are glutathionylated, with mutations of these three cysteines to serines attenuating microtubule dynamics but buffering diamide-induced decrease in microtubule dynamics. In addition, glutaredoxin 1 (Grx1) deglutathionylates EB1, and Grx1 depletion suppresses microtubule dynamics and leads to defects in cell division orientation and cell migration, suggesting a critical role of Grx1-mediated deglutathionylation in maintaining EB1 activity. Collectively, these data reveal that EB1 glutathionylation is an important protective mechanism for the regulation of microtubule dynamics and microtubule-based cellular activities.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (31701216, 31771542, 31900502), and the Natural Science Foundation of Shandong Province (ZR2017MC008). We thank Dr. Hongbo Luo for discussion.

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  1. Contributed equally to this work

Affiliations

  1. Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Sciences, Collaborative Innovation Center of Cell Biology in Universities of Shandong, Institute of Biomedical Sciences, Shandong Normal University, Jinan, 250014, China

    Miao Chen, Tao Zhong, Peng Zhou, Huixian Ma, Jingrui Li, Jun Zhou, Songbo Xie & Min Liu

  2. State Key Laboratory of Medicinal Chemical Biology, Key Laboratory of Bioactive Materials of the Ministry of Education, Tianjin Key Laboratory of Protein Science, College of Life Sciences, Nankai University, Tianjin, 300071, China

    Jian Wang, Dengwen Li & Jun Zhou

  3. Department of Translational Medicine Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China

    Yang Yang

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  1. Miao Chen
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  2. Jian Wang
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  3. Yang Yang
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  4. Tao Zhong
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  5. Peng Zhou
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  7. Jingrui Li
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  8. Dengwen Li
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  9. Jun Zhou
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  10. Songbo Xie
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  11. Min Liu
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Correspondence to Songbo Xie or Min Liu.

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Chen, M., Wang, J., Yang, Y. et al. Redox-dependent regulation of end-binding protein 1 activity by glutathionylation. Sci. China Life Sci. 64, 575–583 (2021). https://doi.org/10.1007/s11427-020-1765-6

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  • DOI: https://doi.org/10.1007/s11427-020-1765-6

Keywords

  • glutathionylation
  • end-binding protein 1
  • microtubule dynamics
  • cell division orientation
  • glutaredoxin 1