Cholesterol Modification of Smoothened Is Required for Hedgehog Signaling

Xu Xiao; Jing Jie Tang; Chao Peng; Yan Wang; Lin Fu; Zhi Ping Qiu; Yue Xiong; Lian Fang Yang; Hai Wei Cui; Xiao Long He; Lei Yin; Wei Qi; Catherine C.L. Wong; Yun Zhao; Bo Liang Li; Wen Wei Qiu; Bao Liang Song

doi:10.1016/j.molcel.2017.02.015

Cholesterol Modification of Smoothened Is Required for Hedgehog Signaling

Xu Xiao, Jing Jie Tang, Chao Peng, Yan Wang, Lin Fu, Zhi Ping Qiu, Yue Xiong, Lian Fang Yang, Hai Wei Cui, Xiao Long He, Lei Yin, Wei Qi, Catherine C.L. Wong, Yun Zhao, Bo Liang Li, Wen Wei Qiu, Bao Liang Song

Physiology

Research output: Contribution to journal › Article › peer-review

186 Scopus citations

Abstract

Hedgehog (Hh) has been known as the only cholesterol-modified morphogen playing pivotal roles in development and tumorigenesis. A major unsolved question is how Hh signaling regulates the activity of Smoothened (SMO). Here, we performed an unbiased biochemical screen and identified that SMO was covalently modified by cholesterol on the Asp95 (D95) residue through an ester bond. This modification was inhibited by Patched-1 (Ptch1) but enhanced by Hh. The SMO(D95N) mutation, which could not be cholesterol modified, was refractory to Hh-stimulated ciliary localization and failed to activate downstream signaling. Furthermore, homozygous Smo^D99N/D99N (the equivalent residue in mouse) knockin mice were embryonic lethal with severe cardiac defects, phenocopying the Smo^−/− mice. Together, the results of our study suggest that Hh signaling transduces to SMO through modulating its cholesterylation and provides a therapeutic opportunity to treat Hh-pathway-related cancers by targeting SMO cholesterylation.

Original language	English
Pages (from-to)	154-162.e10
Journal	Molecular Cell
Volume	66
Issue number	1
DOIs	https://doi.org/10.1016/j.molcel.2017.02.015
State	Published - Apr 6 2017

Bibliographical note

Publisher Copyright:
© 2017 Elsevier Inc.

Funding

We thank Jie Xu, Yu-Xiu Qu, and Hong-Hua Miao for technical assistance; we thank Drs. Bin Zhou, Qiang-Qiang Han, and Jie Luo for helpful discussion. This work was supported by the Ministry of Science and Technology of China (2016YFA0500100), the National Natural Science Foundation of China (31430044, 31230020, and 81270155), the 111 Project of Ministry of Education of China (B16036), the National Key Technology R&D Program (2015BAK45B00), the “Strategic Priority Research Program” of the Chinese Academy of Sciences (XDB19020101), the Shanghai Academic/Technology Research Leader Program (17XD1404100) and a SA-SIBS Discovery Innovation Grant.

Funders	Funder number
111 Project of Ministry of Education of China	B16036
SA-SIBS
National Natural Science Foundation of China (NSFC)	31430044, 31230020, 81270155
Chinese Academy of Sciences	XDB19020101
Ministry of Science and Technology of the People's Republic of China	2016YFA0500100
National Key Basic Research and Development Program of China	2015BAK45B00
Program of Shanghai Academic Research Leader	17XD1404100

Keywords

cholesterol
cholesterylation
ciliary localization
click chemistry
cysteine-rich domain
embryonic lethal
Hedgehog
Patched-1
primary cilia
Smoothened

ASJC Scopus subject areas

Molecular Biology
Cell Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.molcel.2017.02.015

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title = "Cholesterol Modification of Smoothened Is Required for Hedgehog Signaling",

abstract = "Hedgehog (Hh) has been known as the only cholesterol-modified morphogen playing pivotal roles in development and tumorigenesis. A major unsolved question is how Hh signaling regulates the activity of Smoothened (SMO). Here, we performed an unbiased biochemical screen and identified that SMO was covalently modified by cholesterol on the Asp95 (D95) residue through an ester bond. This modification was inhibited by Patched-1 (Ptch1) but enhanced by Hh. The SMO(D95N) mutation, which could not be cholesterol modified, was refractory to Hh-stimulated ciliary localization and failed to activate downstream signaling. Furthermore, homozygous SmoD99N/D99N (the equivalent residue in mouse) knockin mice were embryonic lethal with severe cardiac defects, phenocopying the Smo−/− mice. Together, the results of our study suggest that Hh signaling transduces to SMO through modulating its cholesterylation and provides a therapeutic opportunity to treat Hh-pathway-related cancers by targeting SMO cholesterylation.",

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language = "English",

volume = "66",

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AU - Qiu, Zhi Ping

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AU - Yang, Lian Fang

AU - Cui, Hai Wei

AU - He, Xiao Long

AU - Yin, Lei

AU - Qi, Wei

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AU - Li, Bo Liang

AU - Qiu, Wen Wei

AU - Song, Bao Liang

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AB - Hedgehog (Hh) has been known as the only cholesterol-modified morphogen playing pivotal roles in development and tumorigenesis. A major unsolved question is how Hh signaling regulates the activity of Smoothened (SMO). Here, we performed an unbiased biochemical screen and identified that SMO was covalently modified by cholesterol on the Asp95 (D95) residue through an ester bond. This modification was inhibited by Patched-1 (Ptch1) but enhanced by Hh. The SMO(D95N) mutation, which could not be cholesterol modified, was refractory to Hh-stimulated ciliary localization and failed to activate downstream signaling. Furthermore, homozygous SmoD99N/D99N (the equivalent residue in mouse) knockin mice were embryonic lethal with severe cardiac defects, phenocopying the Smo−/− mice. Together, the results of our study suggest that Hh signaling transduces to SMO through modulating its cholesterylation and provides a therapeutic opportunity to treat Hh-pathway-related cancers by targeting SMO cholesterylation.

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Cholesterol Modification of Smoothened Is Required for Hedgehog Signaling

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

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