C-mannosylation of R-spondin3 regulates its secretion and activity of Wnt/β-catenin signaling in cells

Miho Fujiwara; Shintaro Kato; Yuki Niwa; Takehiro Suzuki; Miyu Tsuchiya; Yukiko Sasazawa; Naoshi Dohmae; Siro Simizu

doi:10.1002/1873-3468.12274

C-mannosylation of R-spondin3 regulates its secretion and activity of Wnt/β-catenin signaling in cells

Miho Fujiwara, Shintaro Kato, Yuki Niwa, Takehiro Suzuki, Miyu Tsuchiya, Yukiko Sasazawa, Naoshi Dohmae, Siro Simizu

Department of Applied Chemistry

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

33 Citations (Scopus)

Abstract

R-spondin3 (Rspo3) is a secreted protein, which acts as an agonist of canonical Wnt/β-catenin signaling that plays an important role in embryonic development and homeostasis. In this study, we focused on C-mannosylation, a unique type of glycosylation, of human Rspo3. Rspo3 has two putative C-mannosylation sites at Trp¹⁵³ and Trp¹⁵⁶; however, it had been unclear whether these sites are C-mannosylated or not. We demonstrated that Rspo3 was C-mannosylated at both Trp¹⁵³ and Trp¹⁵⁶ by mass spectrometry. Using C-mannosylation-defective Rspo3 mutant-overexpressing cell lines, we found that C-mannosylation of Rspo3 promotes its secretion and activates Wnt/β-catenin signaling.

Original language	English
Pages (from-to)	2639-2649
Number of pages	11
Journal	FEBS Letters
DOIs	https://doi.org/10.1002/1873-3468.12274
Publication status	Published - 2016 Aug 1

Keywords

C-mannosylation
R-spondin3
Wnt/β-catenin signaling
glycobiology
mass spectrometry
signal transduction

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1002/1873-3468.12274

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title = "C-mannosylation of R-spondin3 regulates its secretion and activity of Wnt/β-catenin signaling in cells",

abstract = "R-spondin3 (Rspo3) is a secreted protein, which acts as an agonist of canonical Wnt/β-catenin signaling that plays an important role in embryonic development and homeostasis. In this study, we focused on C-mannosylation, a unique type of glycosylation, of human Rspo3. Rspo3 has two putative C-mannosylation sites at Trp153 and Trp156; however, it had been unclear whether these sites are C-mannosylated or not. We demonstrated that Rspo3 was C-mannosylated at both Trp153 and Trp156 by mass spectrometry. Using C-mannosylation-defective Rspo3 mutant-overexpressing cell lines, we found that C-mannosylation of Rspo3 promotes its secretion and activates Wnt/β-catenin signaling.",

keywords = "C-mannosylation, R-spondin3, Wnt/β-catenin signaling, glycobiology, mass spectrometry, signal transduction",

author = "Miho Fujiwara and Shintaro Kato and Yuki Niwa and Takehiro Suzuki and Miyu Tsuchiya and Yukiko Sasazawa and Naoshi Dohmae and Siro Simizu",

note = "Funding Information: This work was supported by the Grants-in-Aid for Scientific Research (B) (24310167), for JSPS Fellows (254256), and a Grant-in-Aid for Young Scientists (B) (26750373). Publisher Copyright: {\textcopyright} 2016 Federation of European Biochemical Societies",

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doi = "10.1002/1873-3468.12274",

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T1 - C-mannosylation of R-spondin3 regulates its secretion and activity of Wnt/β-catenin signaling in cells

AU - Fujiwara, Miho

AU - Kato, Shintaro

AU - Niwa, Yuki

AU - Suzuki, Takehiro

AU - Tsuchiya, Miyu

AU - Sasazawa, Yukiko

AU - Dohmae, Naoshi

AU - Simizu, Siro

N1 - Funding Information: This work was supported by the Grants-in-Aid for Scientific Research (B) (24310167), for JSPS Fellows (254256), and a Grant-in-Aid for Young Scientists (B) (26750373). Publisher Copyright: © 2016 Federation of European Biochemical Societies

PY - 2016/8/1

Y1 - 2016/8/1

N2 - R-spondin3 (Rspo3) is a secreted protein, which acts as an agonist of canonical Wnt/β-catenin signaling that plays an important role in embryonic development and homeostasis. In this study, we focused on C-mannosylation, a unique type of glycosylation, of human Rspo3. Rspo3 has two putative C-mannosylation sites at Trp153 and Trp156; however, it had been unclear whether these sites are C-mannosylated or not. We demonstrated that Rspo3 was C-mannosylated at both Trp153 and Trp156 by mass spectrometry. Using C-mannosylation-defective Rspo3 mutant-overexpressing cell lines, we found that C-mannosylation of Rspo3 promotes its secretion and activates Wnt/β-catenin signaling.

AB - R-spondin3 (Rspo3) is a secreted protein, which acts as an agonist of canonical Wnt/β-catenin signaling that plays an important role in embryonic development and homeostasis. In this study, we focused on C-mannosylation, a unique type of glycosylation, of human Rspo3. Rspo3 has two putative C-mannosylation sites at Trp153 and Trp156; however, it had been unclear whether these sites are C-mannosylated or not. We demonstrated that Rspo3 was C-mannosylated at both Trp153 and Trp156 by mass spectrometry. Using C-mannosylation-defective Rspo3 mutant-overexpressing cell lines, we found that C-mannosylation of Rspo3 promotes its secretion and activates Wnt/β-catenin signaling.

KW - C-mannosylation

KW - R-spondin3

KW - Wnt/β-catenin signaling

KW - glycobiology

KW - mass spectrometry

KW - signal transduction

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C-mannosylation of R-spondin3 regulates its secretion and activity of Wnt/β-catenin signaling in cells

Abstract

Keywords

ASJC Scopus subject areas

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