C‑mannosylation of R‑spondin2 activates Wnt/β‑catenin signaling and migration activity in human tumor cells

Hayato Mizuta, Kenta Kuga, Takehiro Suzuki, Yuki Niwa, Naoshi Dohmae, Siro Simizu

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

R-spondin2 (Rspo2), one of the four members of the R-spondin family of proteins, has agonistic activity in the Wnt/β-catenin signaling pathway, and it is associated with normal development, as well as disease, such as cancer. The present study focused on the C-mannosylation of Rspo2, which is a novel and unique type of glycosylation that occurs via a C-C linkage between the tryptophan residue and an α-mannose. Although Rspo2 has two putative C-mannosylation sites at residues Trp 150 and Trp 153 , it had not been reported to date whether these sites are C-mannosylated. Firstly, results from mass spectrometry demonstrated that Rspo2 was C-mannosylated at the Trp 150 and Trp 153 residues. Notably, while this C-mannosylation of Rspo2 resulted in increased extracellular secretion in human fibrosarcoma HT1080 cells, in other human tumor cell lines it inhibited secretion. However, C-mannosylation had consistent effects on the activation of Wnt/β-catenin signaling in PANC1 and MDA-MB-231 cells, as well as HT1080 cells. Furthermore, overexpression of wild-type Rspo2 significantly increased the migratory ability of A549 and HT1080 cells, whereas overexpression of a C-mannosylation-defective mutant enhanced migration to a lesser degree. These results suggested that C-mannosylation of Rspo2 may promote cancer progression and that the inhibition of C-mannosylation may serve as a potential novel therapeutic approach for cancer therapy.

Original languageEnglish
Pages (from-to)2127-2138
Number of pages12
JournalInternational journal of oncology
Volume54
Issue number6
DOIs
Publication statusPublished - 2019 Jun 1

Fingerprint

Catenins
Human Activities
Neoplasms
Wnt Signaling Pathway
Fibrosarcoma
Mannose
Tumor Cell Line
Glycosylation
Tryptophan
Mass Spectrometry
Therapeutics
Proteins

Keywords

  • C-mannosylation
  • Cell migration
  • Intracellular trafficking
  • R-spondin2
  • Wnt/β-catenin signaling

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

C‑mannosylation of R‑spondin2 activates Wnt/β‑catenin signaling and migration activity in human tumor cells. / Mizuta, Hayato; Kuga, Kenta; Suzuki, Takehiro; Niwa, Yuki; Dohmae, Naoshi; Simizu, Siro.

In: International journal of oncology, Vol. 54, No. 6, 01.06.2019, p. 2127-2138.

Research output: Contribution to journalArticle

Mizuta, Hayato ; Kuga, Kenta ; Suzuki, Takehiro ; Niwa, Yuki ; Dohmae, Naoshi ; Simizu, Siro. / C‑mannosylation of R‑spondin2 activates Wnt/β‑catenin signaling and migration activity in human tumor cells. In: International journal of oncology. 2019 ; Vol. 54, No. 6. pp. 2127-2138.
@article{5187dc200c2d41298f214eadb1c7b8e3,
title = "C‑mannosylation of R‑spondin2 activates Wnt/β‑catenin signaling and migration activity in human tumor cells",
abstract = "R-spondin2 (Rspo2), one of the four members of the R-spondin family of proteins, has agonistic activity in the Wnt/β-catenin signaling pathway, and it is associated with normal development, as well as disease, such as cancer. The present study focused on the C-mannosylation of Rspo2, which is a novel and unique type of glycosylation that occurs via a C-C linkage between the tryptophan residue and an α-mannose. Although Rspo2 has two putative C-mannosylation sites at residues Trp 150 and Trp 153 , it had not been reported to date whether these sites are C-mannosylated. Firstly, results from mass spectrometry demonstrated that Rspo2 was C-mannosylated at the Trp 150 and Trp 153 residues. Notably, while this C-mannosylation of Rspo2 resulted in increased extracellular secretion in human fibrosarcoma HT1080 cells, in other human tumor cell lines it inhibited secretion. However, C-mannosylation had consistent effects on the activation of Wnt/β-catenin signaling in PANC1 and MDA-MB-231 cells, as well as HT1080 cells. Furthermore, overexpression of wild-type Rspo2 significantly increased the migratory ability of A549 and HT1080 cells, whereas overexpression of a C-mannosylation-defective mutant enhanced migration to a lesser degree. These results suggested that C-mannosylation of Rspo2 may promote cancer progression and that the inhibition of C-mannosylation may serve as a potential novel therapeutic approach for cancer therapy.",
keywords = "C-mannosylation, Cell migration, Intracellular trafficking, R-spondin2, Wnt/β-catenin signaling",
author = "Hayato Mizuta and Kenta Kuga and Takehiro Suzuki and Yuki Niwa and Naoshi Dohmae and Siro Simizu",
year = "2019",
month = "6",
day = "1",
doi = "10.3892/ijo.2019.4767",
language = "English",
volume = "54",
pages = "2127--2138",
journal = "International Journal of Oncology",
issn = "1019-6439",
publisher = "Spandidos Publications",
number = "6",

}

TY - JOUR

T1 - C‑mannosylation of R‑spondin2 activates Wnt/β‑catenin signaling and migration activity in human tumor cells

AU - Mizuta, Hayato

AU - Kuga, Kenta

AU - Suzuki, Takehiro

AU - Niwa, Yuki

AU - Dohmae, Naoshi

AU - Simizu, Siro

PY - 2019/6/1

Y1 - 2019/6/1

N2 - R-spondin2 (Rspo2), one of the four members of the R-spondin family of proteins, has agonistic activity in the Wnt/β-catenin signaling pathway, and it is associated with normal development, as well as disease, such as cancer. The present study focused on the C-mannosylation of Rspo2, which is a novel and unique type of glycosylation that occurs via a C-C linkage between the tryptophan residue and an α-mannose. Although Rspo2 has two putative C-mannosylation sites at residues Trp 150 and Trp 153 , it had not been reported to date whether these sites are C-mannosylated. Firstly, results from mass spectrometry demonstrated that Rspo2 was C-mannosylated at the Trp 150 and Trp 153 residues. Notably, while this C-mannosylation of Rspo2 resulted in increased extracellular secretion in human fibrosarcoma HT1080 cells, in other human tumor cell lines it inhibited secretion. However, C-mannosylation had consistent effects on the activation of Wnt/β-catenin signaling in PANC1 and MDA-MB-231 cells, as well as HT1080 cells. Furthermore, overexpression of wild-type Rspo2 significantly increased the migratory ability of A549 and HT1080 cells, whereas overexpression of a C-mannosylation-defective mutant enhanced migration to a lesser degree. These results suggested that C-mannosylation of Rspo2 may promote cancer progression and that the inhibition of C-mannosylation may serve as a potential novel therapeutic approach for cancer therapy.

AB - R-spondin2 (Rspo2), one of the four members of the R-spondin family of proteins, has agonistic activity in the Wnt/β-catenin signaling pathway, and it is associated with normal development, as well as disease, such as cancer. The present study focused on the C-mannosylation of Rspo2, which is a novel and unique type of glycosylation that occurs via a C-C linkage between the tryptophan residue and an α-mannose. Although Rspo2 has two putative C-mannosylation sites at residues Trp 150 and Trp 153 , it had not been reported to date whether these sites are C-mannosylated. Firstly, results from mass spectrometry demonstrated that Rspo2 was C-mannosylated at the Trp 150 and Trp 153 residues. Notably, while this C-mannosylation of Rspo2 resulted in increased extracellular secretion in human fibrosarcoma HT1080 cells, in other human tumor cell lines it inhibited secretion. However, C-mannosylation had consistent effects on the activation of Wnt/β-catenin signaling in PANC1 and MDA-MB-231 cells, as well as HT1080 cells. Furthermore, overexpression of wild-type Rspo2 significantly increased the migratory ability of A549 and HT1080 cells, whereas overexpression of a C-mannosylation-defective mutant enhanced migration to a lesser degree. These results suggested that C-mannosylation of Rspo2 may promote cancer progression and that the inhibition of C-mannosylation may serve as a potential novel therapeutic approach for cancer therapy.

KW - C-mannosylation

KW - Cell migration

KW - Intracellular trafficking

KW - R-spondin2

KW - Wnt/β-catenin signaling

UR - http://www.scopus.com/inward/record.url?scp=85064860605&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85064860605&partnerID=8YFLogxK

U2 - 10.3892/ijo.2019.4767

DO - 10.3892/ijo.2019.4767

M3 - Article

C2 - 30942431

AN - SCOPUS:85064860605

VL - 54

SP - 2127

EP - 2138

JO - International Journal of Oncology

JF - International Journal of Oncology

SN - 1019-6439

IS - 6

ER -