Cholesterylation of Smoothened is a calcium-accelerated autoreaction involving an intramolecular ester intermediate

Ao Hu; Jing Zan Zhang; Jie Wang; Chen Chen Li; Meng Yuan; Gang Deng; Zi Cun Lin; Zhi Ping Qiu; Hu Yue Liu; Xian Wei Wang; Peng Cheng Wei; Xiao He; Xiaolu Zhao; Wen Wei Qiu; Bao Liang Song

doi:10.1038/s41422-022-00622-0

Cholesterylation of Smoothened is a calcium-accelerated autoreaction involving an intramolecular ester intermediate

Ao Hu, Jing Zan Zhang, Jie Wang, Chen Chen Li, Meng Yuan, Gang Deng, Zi Cun Lin, Zhi Ping Qiu, Hu Yue Liu, Xian Wei Wang, Peng Cheng Wei, Xiao He, Xiaolu Zhao, Wen Wei Qiu, Bao Liang Song

School of Chemistry and Molecular Engineering

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

27 Scopus citations

Abstract

Hedgehog (Hh) is a morphogen that binds to its receptor Patched 1 and activates Smoothened (SMO), thereby governing embryonic development and postnatal tissue homeostasis. Cholesterol can bind and covalently conjugate to the luminal cysteine-rich domain (CRD) of human SMO at the D95 residue (D99 in mouse). The reaction mechanism and biological function of SMO cholesterylation have not been elucidated. Here, we show that the SMO-CRD undergoes auto-cholesterylation which is boosted by calcium and involves an intramolecular ester intermediate. In cells, Hh stimulation elevates local calcium concentration in the SMO-localized endosomes through store-operated calcium entry. In addition, we identify the signaling-incompetent SMO D95E mutation, and the D95E mutant SMO can bind cholesterol but cannot be modified or activated by cholesterol. The homozygous Smo^D99E/D99E knockin mice are embryonic lethal with severe developmental delay, demonstrating that cholesterylation of CRD is required for full-length SMO activation. Our work reveals the unique autocatalytic mechanism of SMO cholesterylation and an unprecedented role of calcium in Hh signaling.

Original language	English
Pages (from-to)	288-301
Number of pages	14
Journal	Cell Research
Volume	32
Issue number	3
DOIs	https://doi.org/10.1038/s41422-022-00622-0
State	Published - Mar 2022

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@article{b9d00fb7beea4acdb753e92a604bc2b5,

title = "Cholesterylation of Smoothened is a calcium-accelerated autoreaction involving an intramolecular ester intermediate",

abstract = "Hedgehog (Hh) is a morphogen that binds to its receptor Patched 1 and activates Smoothened (SMO), thereby governing embryonic development and postnatal tissue homeostasis. Cholesterol can bind and covalently conjugate to the luminal cysteine-rich domain (CRD) of human SMO at the D95 residue (D99 in mouse). The reaction mechanism and biological function of SMO cholesterylation have not been elucidated. Here, we show that the SMO-CRD undergoes auto-cholesterylation which is boosted by calcium and involves an intramolecular ester intermediate. In cells, Hh stimulation elevates local calcium concentration in the SMO-localized endosomes through store-operated calcium entry. In addition, we identify the signaling-incompetent SMO D95E mutation, and the D95E mutant SMO can bind cholesterol but cannot be modified or activated by cholesterol. The homozygous SmoD99E/D99E knockin mice are embryonic lethal with severe developmental delay, demonstrating that cholesterylation of CRD is required for full-length SMO activation. Our work reveals the unique autocatalytic mechanism of SMO cholesterylation and an unprecedented role of calcium in Hh signaling.",

author = "Ao Hu and Zhang, \{Jing Zan\} and Jie Wang and Li, \{Chen Chen\} and Meng Yuan and Gang Deng and Lin, \{Zi Cun\} and Qiu, \{Zhi Ping\} and Liu, \{Hu Yue\} and Wang, \{Xian Wei\} and Wei, \{Peng Cheng\} and Xiao He and Xiaolu Zhao and Qiu, \{Wen Wei\} and Song, \{Bao Liang\}",

note = "Publisher Copyright: {\textcopyright} 2022, The Author(s) under exclusive licence to Center for Excellence in Molecular Cell Science, CAS.",

year = "2022",

month = mar,

doi = "10.1038/s41422-022-00622-0",

language = "英语",

volume = "32",

pages = "288--301",

journal = "Cell Research",

issn = "1001-0602",

publisher = "Springer Nature",

number = "3",

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T1 - Cholesterylation of Smoothened is a calcium-accelerated autoreaction involving an intramolecular ester intermediate

AU - Hu, Ao

AU - Zhang, Jing Zan

AU - Wang, Jie

AU - Li, Chen Chen

AU - Yuan, Meng

AU - Deng, Gang

AU - Lin, Zi Cun

AU - Qiu, Zhi Ping

AU - Liu, Hu Yue

AU - Wang, Xian Wei

AU - Wei, Peng Cheng

AU - He, Xiao

AU - Zhao, Xiaolu

AU - Qiu, Wen Wei

AU - Song, Bao Liang

PY - 2022/3

Y1 - 2022/3

N2 - Hedgehog (Hh) is a morphogen that binds to its receptor Patched 1 and activates Smoothened (SMO), thereby governing embryonic development and postnatal tissue homeostasis. Cholesterol can bind and covalently conjugate to the luminal cysteine-rich domain (CRD) of human SMO at the D95 residue (D99 in mouse). The reaction mechanism and biological function of SMO cholesterylation have not been elucidated. Here, we show that the SMO-CRD undergoes auto-cholesterylation which is boosted by calcium and involves an intramolecular ester intermediate. In cells, Hh stimulation elevates local calcium concentration in the SMO-localized endosomes through store-operated calcium entry. In addition, we identify the signaling-incompetent SMO D95E mutation, and the D95E mutant SMO can bind cholesterol but cannot be modified or activated by cholesterol. The homozygous SmoD99E/D99E knockin mice are embryonic lethal with severe developmental delay, demonstrating that cholesterylation of CRD is required for full-length SMO activation. Our work reveals the unique autocatalytic mechanism of SMO cholesterylation and an unprecedented role of calcium in Hh signaling.

AB - Hedgehog (Hh) is a morphogen that binds to its receptor Patched 1 and activates Smoothened (SMO), thereby governing embryonic development and postnatal tissue homeostasis. Cholesterol can bind and covalently conjugate to the luminal cysteine-rich domain (CRD) of human SMO at the D95 residue (D99 in mouse). The reaction mechanism and biological function of SMO cholesterylation have not been elucidated. Here, we show that the SMO-CRD undergoes auto-cholesterylation which is boosted by calcium and involves an intramolecular ester intermediate. In cells, Hh stimulation elevates local calcium concentration in the SMO-localized endosomes through store-operated calcium entry. In addition, we identify the signaling-incompetent SMO D95E mutation, and the D95E mutant SMO can bind cholesterol but cannot be modified or activated by cholesterol. The homozygous SmoD99E/D99E knockin mice are embryonic lethal with severe developmental delay, demonstrating that cholesterylation of CRD is required for full-length SMO activation. Our work reveals the unique autocatalytic mechanism of SMO cholesterylation and an unprecedented role of calcium in Hh signaling.

UR - https://www.scopus.com/pages/publications/85124276843

U2 - 10.1038/s41422-022-00622-0

DO - 10.1038/s41422-022-00622-0

M3 - 文章

C2 - 35121857

AN - SCOPUS:85124276843

SN - 1001-0602

VL - 32

SP - 288

EP - 301

JO - Cell Research

JF - Cell Research

IS - 3

ER -

Cholesterylation of Smoothened is a calcium-accelerated autoreaction involving an intramolecular ester intermediate

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