The Innovative Role of Nuclear Receptor Interaction Protein in Orchestrating Invadosome Formation for Myoblast Fusion
ABSTRACT Background Nuclear receptor interaction protein (NRIP) is versatile and engages with various proteins to execute its diverse biological function. NRIP deficiency was reported to cause small myofibre size in adult muscle regeneration, indicating a crucial role of NRIP in myoblast fusion. Met...
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Wiley
2024-12-01
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| Series: | Journal of Cachexia, Sarcopenia and Muscle |
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| Online Access: | https://doi.org/10.1002/jcsm.13598 |
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| author | Hsin‐Hsiung Chen Chia‐Yang Lin Ya‐Ju Han Yun‐Hsin Huang Yi‐Hsiang Liu Wan‐En Hsu Li‐Kai Tsai Hsing‐Jung Lai Yeou‐Ping Tsao Hsiang‐Po Huang Show‐Li Chen |
| author_facet | Hsin‐Hsiung Chen Chia‐Yang Lin Ya‐Ju Han Yun‐Hsin Huang Yi‐Hsiang Liu Wan‐En Hsu Li‐Kai Tsai Hsing‐Jung Lai Yeou‐Ping Tsao Hsiang‐Po Huang Show‐Li Chen |
| author_sort | Hsin‐Hsiung Chen |
| collection | DOAJ |
| description | ABSTRACT Background Nuclear receptor interaction protein (NRIP) is versatile and engages with various proteins to execute its diverse biological function. NRIP deficiency was reported to cause small myofibre size in adult muscle regeneration, indicating a crucial role of NRIP in myoblast fusion. Methods The colocalization and interaction of NRIP with actin were investigated by immunofluorescence and immunoprecipitation assay, respectively. The participation of NRIP in myoblast fusion was demonstrated by cell fusion assay and time‐lapse microscopy. The NRIP mutants were generated for mechanism study in NRIP‐null C2C12 (termed KO19) cells and muscle‐specific NRIP knockout (NRIP cKO) mice. A GEO profile database was used to analyse NRIP expression in Duchenne muscular dystrophy (DMD) patients. Results In this study, we found that NRIP directly and reciprocally interacted with actin both in vitro and in cells. Immunofluorescence microscopy showed that the endogenous NRIP colocalized with components of invadosome, such as actin, Tks5, and cortactin, at the tips of cells during C2C12 differentiation. The KO19 cells were generated and exhibited a significant deficit in myoblast fusion compared with wild‐type C2C12 cells (3.16% vs. 33.67%, p < 0.005). Overexpressed NRIP in KO19 cells could rescue myotube formation compared with control (3.37% vs. 1.00%, p < 0.01). We further confirmed that NRIP directly participated in cell fusion by using a cell–cell fusion assay. We investigated the mechanism of invadosome formation for myoblast fusion, which depends on NRIP–actin interaction, by analysing NRIP mutants in NRIP‐null cells. Loss of actin‐binding of NRIP reduced invadosome (enrichment ratio, 1.00 vs. 2.54, p < 0.01) and myotube formation (21.82% vs. 35.71%, p < 0.05) in KO19 cells and forced NRIP expression in KO19 cells and muscle‐specific NRIP knockout (NRIP cKO) mice increased myofibre size compared with controls (over 1500 μm2, 61.01% vs. 20.57%, p < 0.001). We also found that the NRIP mRNA level was decreased in DMD patients compared with healthy controls (18 072 vs. 28 289, p < 0.001, N = 10 for both groups). Conclusions NRIP is a novel actin‐binding protein for invadosome formation to induce myoblast fusion. |
| format | Article |
| id | doaj-art-50fa6166eeac479199cfac2eb4e0c6f6 |
| institution | OA Journals |
| issn | 2190-5991 2190-6009 |
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| publishDate | 2024-12-01 |
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| series | Journal of Cachexia, Sarcopenia and Muscle |
| spelling | doaj-art-50fa6166eeac479199cfac2eb4e0c6f62025-08-20T02:34:20ZengWileyJournal of Cachexia, Sarcopenia and Muscle2190-59912190-60092024-12-011562559257310.1002/jcsm.13598The Innovative Role of Nuclear Receptor Interaction Protein in Orchestrating Invadosome Formation for Myoblast FusionHsin‐Hsiung Chen0Chia‐Yang Lin1Ya‐Ju Han2Yun‐Hsin Huang3Yi‐Hsiang Liu4Wan‐En Hsu5Li‐Kai Tsai6Hsing‐Jung Lai7Yeou‐Ping Tsao8Hsiang‐Po Huang9Show‐Li Chen10Graduate Institute of Microbiology, College of Medicine National Taiwan University Taipei TaiwanGraduate Institute of Microbiology, College of Medicine National Taiwan University Taipei TaiwanGraduate Institute of Microbiology, College of Medicine National Taiwan University Taipei TaiwanGraduate Institute of Microbiology, College of Medicine National Taiwan University Taipei TaiwanGraduate Institute of Microbiology, College of Medicine National Taiwan University Taipei TaiwanGraduate Institute of Microbiology, College of Medicine National Taiwan University Taipei TaiwanDepartment of Neurology National Taiwan University Hospital Taipei TaiwanDepartment of Neurology National Taiwan University Hospital Taipei TaiwanDepartment of Ophthalmology Mackay Memorial Hospital Taipei TaiwanGraduate Institute of Medical Genomics and Proteomics, College of Medicine National Taiwan University Taipei TaiwanGraduate Institute of Microbiology, College of Medicine National Taiwan University Taipei TaiwanABSTRACT Background Nuclear receptor interaction protein (NRIP) is versatile and engages with various proteins to execute its diverse biological function. NRIP deficiency was reported to cause small myofibre size in adult muscle regeneration, indicating a crucial role of NRIP in myoblast fusion. Methods The colocalization and interaction of NRIP with actin were investigated by immunofluorescence and immunoprecipitation assay, respectively. The participation of NRIP in myoblast fusion was demonstrated by cell fusion assay and time‐lapse microscopy. The NRIP mutants were generated for mechanism study in NRIP‐null C2C12 (termed KO19) cells and muscle‐specific NRIP knockout (NRIP cKO) mice. A GEO profile database was used to analyse NRIP expression in Duchenne muscular dystrophy (DMD) patients. Results In this study, we found that NRIP directly and reciprocally interacted with actin both in vitro and in cells. Immunofluorescence microscopy showed that the endogenous NRIP colocalized with components of invadosome, such as actin, Tks5, and cortactin, at the tips of cells during C2C12 differentiation. The KO19 cells were generated and exhibited a significant deficit in myoblast fusion compared with wild‐type C2C12 cells (3.16% vs. 33.67%, p < 0.005). Overexpressed NRIP in KO19 cells could rescue myotube formation compared with control (3.37% vs. 1.00%, p < 0.01). We further confirmed that NRIP directly participated in cell fusion by using a cell–cell fusion assay. We investigated the mechanism of invadosome formation for myoblast fusion, which depends on NRIP–actin interaction, by analysing NRIP mutants in NRIP‐null cells. Loss of actin‐binding of NRIP reduced invadosome (enrichment ratio, 1.00 vs. 2.54, p < 0.01) and myotube formation (21.82% vs. 35.71%, p < 0.05) in KO19 cells and forced NRIP expression in KO19 cells and muscle‐specific NRIP knockout (NRIP cKO) mice increased myofibre size compared with controls (over 1500 μm2, 61.01% vs. 20.57%, p < 0.001). We also found that the NRIP mRNA level was decreased in DMD patients compared with healthy controls (18 072 vs. 28 289, p < 0.001, N = 10 for both groups). Conclusions NRIP is a novel actin‐binding protein for invadosome formation to induce myoblast fusion.https://doi.org/10.1002/jcsm.13598actininvadosomeIQ domainmyoblast fusionNRIPWD40 domain |
| spellingShingle | Hsin‐Hsiung Chen Chia‐Yang Lin Ya‐Ju Han Yun‐Hsin Huang Yi‐Hsiang Liu Wan‐En Hsu Li‐Kai Tsai Hsing‐Jung Lai Yeou‐Ping Tsao Hsiang‐Po Huang Show‐Li Chen The Innovative Role of Nuclear Receptor Interaction Protein in Orchestrating Invadosome Formation for Myoblast Fusion Journal of Cachexia, Sarcopenia and Muscle actin invadosome IQ domain myoblast fusion NRIP WD40 domain |
| title | The Innovative Role of Nuclear Receptor Interaction Protein in Orchestrating Invadosome Formation for Myoblast Fusion |
| title_full | The Innovative Role of Nuclear Receptor Interaction Protein in Orchestrating Invadosome Formation for Myoblast Fusion |
| title_fullStr | The Innovative Role of Nuclear Receptor Interaction Protein in Orchestrating Invadosome Formation for Myoblast Fusion |
| title_full_unstemmed | The Innovative Role of Nuclear Receptor Interaction Protein in Orchestrating Invadosome Formation for Myoblast Fusion |
| title_short | The Innovative Role of Nuclear Receptor Interaction Protein in Orchestrating Invadosome Formation for Myoblast Fusion |
| title_sort | innovative role of nuclear receptor interaction protein in orchestrating invadosome formation for myoblast fusion |
| topic | actin invadosome IQ domain myoblast fusion NRIP WD40 domain |
| url | https://doi.org/10.1002/jcsm.13598 |
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