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Title: 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. 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:
; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
FOREIGN
OSTI Identifier:
1601858
Resource Type:
Journal Article
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

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. 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, and Chen, Zhucheng. Mon . "Structure and regulation of human epithelial cell transforming 2 protein". United States. doi:10.1073/pnas.1913054117.
@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. 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},
issn = {0027-8424},
number = 2,
volume = 117,
place = {United States},
year = {2019},
month = {12}
}

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