Structure and regulation of human epithelial cell transforming 2 protein
Abstract
Epithelial cell transforming 2 (Ect2) protein activates Rho GTPases and controls cytokinesis and many other cellular processes. Dysregulation of Ect2 is associated with various cancers. Here, we report the crystal structure of human Ect2 and complementary mechanistic analyses. The data show the C-terminal PH domain of Ect2 folds back and blocks the canonical RhoA-binding site at the catalytic center of the DH domain, providing a mechanism of Ect2 autoinhibition. Ect2 is activated by binding of GTP-bound RhoA to the PH domain, which suggests an allosteric mechanism of Ect2 activation and a positive-feedback loop reinforcing RhoA signaling. Furthermore this bimodal RhoA binding of Ect2 is unusual and was confirmed with Förster resonance energy transfer (FRET) and hydrogen–deuterium exchange mass spectrometry (HDX-MS) analyses. Several recurrent cancer-associated mutations map to the catalytic and regulatory interfaces, and dysregulate Ect2 in vitro and in vivo. Together, our findings provide mechanistic insights into Ect2 regulation in normal cells and under disease conditions.
- Authors:
-
- Tsinghua Univ., Beijing (China); Peking Univ., Beijing (China)
- Tsinghua Univ., Beijing (China)
- Tsinghua Univ., Beijing (China); Waigaoqiao Free Trade Zone, Shanghai (China)
- Peking Univ., Beijing (China)
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- National Natural Science Foundation of China (NSFC)
- OSTI Identifier:
- 1601858
- Grant/Contract Number:
- 2019YFA0508900; 2017YFA0102900; 31570731; 31270762; 31630046
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Proceedings of the National Academy of Sciences of the United States of America
- Additional Journal Information:
- Journal Volume: 117; Journal Issue: 2; Journal ID: ISSN 0027-8424
- Publisher:
- National Academy of Sciences
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; cytokinesis; Ect2; RhoA; cancer; biophysics
Citation Formats
Chen, Mengran, Pan, Han, Sun, Lingfei, Shi, Peng, Zhang, Yikan, Li, Le, Huang, Yuxing, Chen, Jianhui, Jiang, Peng, Fang, Xianyang, Wu, Congying, and Chen, Zhucheng. Structure and regulation of human epithelial cell transforming 2 protein. United States: N. p., 2019.
Web. doi:10.1073/pnas.1913054117.
Chen, Mengran, Pan, Han, Sun, Lingfei, Shi, Peng, Zhang, Yikan, Li, Le, Huang, Yuxing, Chen, Jianhui, Jiang, Peng, Fang, Xianyang, Wu, Congying, & Chen, Zhucheng. Structure and regulation of human epithelial cell transforming 2 protein. United States. https://doi.org/10.1073/pnas.1913054117
Chen, Mengran, Pan, Han, Sun, Lingfei, Shi, Peng, Zhang, Yikan, Li, Le, Huang, Yuxing, Chen, Jianhui, Jiang, Peng, Fang, Xianyang, Wu, Congying, and Chen, Zhucheng. Mon .
"Structure and regulation of human epithelial cell transforming 2 protein". United States. https://doi.org/10.1073/pnas.1913054117. https://www.osti.gov/servlets/purl/1601858.
@article{osti_1601858,
title = {Structure and regulation of human epithelial cell transforming 2 protein},
author = {Chen, Mengran and Pan, Han and Sun, Lingfei and Shi, Peng and Zhang, Yikan and Li, Le and Huang, Yuxing and Chen, Jianhui and Jiang, Peng and Fang, Xianyang and Wu, Congying and Chen, Zhucheng},
abstractNote = {Epithelial cell transforming 2 (Ect2) protein activates Rho GTPases and controls cytokinesis and many other cellular processes. Dysregulation of Ect2 is associated with various cancers. Here, we report the crystal structure of human Ect2 and complementary mechanistic analyses. The data show the C-terminal PH domain of Ect2 folds back and blocks the canonical RhoA-binding site at the catalytic center of the DH domain, providing a mechanism of Ect2 autoinhibition. Ect2 is activated by binding of GTP-bound RhoA to the PH domain, which suggests an allosteric mechanism of Ect2 activation and a positive-feedback loop reinforcing RhoA signaling. Furthermore this bimodal RhoA binding of Ect2 is unusual and was confirmed with Förster resonance energy transfer (FRET) and hydrogen–deuterium exchange mass spectrometry (HDX-MS) analyses. Several recurrent cancer-associated mutations map to the catalytic and regulatory interfaces, and dysregulate Ect2 in vitro and in vivo. Together, our findings provide mechanistic insights into Ect2 regulation in normal cells and under disease conditions.},
doi = {10.1073/pnas.1913054117},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 2,
volume = 117,
place = {United States},
year = {2019},
month = {12}
}
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