β-catenin regulates parathyroid hormone/parathyroid hormone-related protein receptor signals and chondrocyte hypertrophy through binding to the intracellular C-terminal region of the receptor.
| Project/Area Number |
21390416
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | The University of Tokyo |
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Principal Investigator |
OGATA Naoshi 東京大学, 大学院・医学系研究科, 講師 (10361495)
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| Co-Investigator(Kenkyū-buntansha) |
CHIKUDA Hirotaka 東京大学, 大学院・医学系研究科, 講師 (30345219)
YANO Fumiko 東京大学, 大学院・医学系研究科, 特任助教 (80529040)
三浦 俊樹 東京大学, 医学部附属病院, 講師 (20376479)
川口 浩 東京大学, 医学部附属病院, 准教授 (40282660)
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| Project Period (FY) |
2009 – 2012
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| Project Status |
Completed (Fiscal Year 2012)
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| Budget Amount *help |
¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
Fiscal Year 2012: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
Fiscal Year 2011: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Fiscal Year 2010: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2009: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
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| Keywords | 細胞・組織 / 骨軟骨代謝 / 副甲状腺ホルモン / 骨粗鬆症 / 骨・軟骨代謝 |
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| Research Abstract |
To investigate the underlying mechanisms of action and functional relevance of ss-catenin in chondrocytes, by examining the role of β-catenin as a novel protein that interacts with the intracellular C-terminal portion of the parathyroid hormone (PTH)/PTH-related protein (PTHrP) receptor type 1 (PTHR-1).The ss-catenin-PTHR-1 binding region was determined with deletion and mutagenesis analyses of the PTHR1 C-terminus, using a mammalian two-hybrid assay. Physical interactions between these 2 molecules were examined with an in situ proximity ligation assay and immunostaining. To assess the effects of gain- and loss-of-function ofssβ-catenin, transfection experiments were performed to induce overexpression of the constitutively active form of ss-catenin (ca-ss-catenin) and to block ss-catenin activity with small interfering RNA, in cells cotransfected with either wild-type PTHR1 or mutant forms (lacking binding to ss-catenin). Activation of the G protein α subunits G(αs) and G(αq) in the ce
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lls was determined by measurement of the intracellular cAMP accumulation and intracellular Ca(2+) concentration, while activation of canonical Wnt pathways was assessed using a TOPflash reporter assay.In differentiated chondrocytes, ss-catenin physically interacted and colocalized with the cell membrane-specific region of PTHR-1 (584-589). Binding of β-catenin to PTHR-1 caused suppression of the G(αs)/cAMP pathway and enhancement of the G(αq)/Ca(2+) pathway, without affecting the canonical Wnt pathway. Inhibition of Col10a1 messenger RNA (mRNA) expression by PTH was restored by overexpression of ca-β-catenin, even after blockade of the canonical Wnt pathway, and Col10a1 mRNA expression was further decreased by knockout of ss-catenin (via the Cre recombinase) in chondrocytes from ss-catenin-floxed mice. Mutagenesis analyses to block the binding of ss-catenin to PTHR1 caused an inhibition of chondrocyte hypertrophy markers.As a conclusion, ss-catenin binds to the PTHR-1 C-tail and switches the downstream signaling pathway from G(αs)/cAMP to G(αq)/Ca(2+), which is a possible mechanism by which chondrocyte hypertrophy may be regulated through the PTH/PTHrP signal independent of thecanonical Wnt pathway. Less
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Report
(5 results)
Research Products
(10 results)
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[Journal Article] β-catenin regulates PTH/PTHrP receptor signals and chondrocyte hypertrophy through binding to its intracellular C-terminal region2013
Author(s)
Yano, F., Saito, T., Ogata, N., Yamazawa, T., Iino, M., Chung, U.I. and Kawaguchi, H
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Journal Title
Arthritis Rheum
Volume: 65
Issue: 2
Pages: 429-435
DOI
Related Report
Peer Reviewed
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