Protein <i>O</i>-Fucosyltransferase 1 Expression Impacts Myogenic C2C12 Cell Commitment via the Notch Signaling Pathway

Vartanian, Audrey Der; Audfray, Aymeric; Jaam, Bilal Al; Janot, Mathilde; Legardinier, Sébastien; Maftah, Abderrahman; Germot, Agnès

doi:10.1128/mcb.00890-14

Cited by 27 publications

(37 citation statements)

References 72 publications

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“…As in Pofut1 knockdown C2C12 cell line (called Po−) [ 44 ], we showed earlier differentiation of Pofut1 cax/cax SCDMs owing to reduced Notch signalling and a significant lowering in PAX7 + /MYOD − progenitor cells in favour of more PAX7 − /MYOD + cells committed to differentiation. Thus, in vitro and ex vivo cell models represented by the C2C12 Po− cell line and Pofut1 cax/cax SCDMs, respectively, gave similar results, highlighting the essential role of POFUT1 in myogenic differentiation through Notch signalling regulation.…”

Section: Discussionsupporting

confidence: 60%

“…Taqman ® primers and probe sets used in this study were as follows: 18S (Hs99999901_s1), Gapdh (Mm99999915_g1), Cdkn1a (Mm00432448_m1), Pofut1 (Mm00475567_m1), Pax7 (Mm03053796_s1), MyoD1 (Mm00440387_m1), Myog (MM00446194_m1), Myf5 (Mm00435125_m1), Hes1 (Mm00468601_m1), Heyl (Mm00516555_m1), Hey1 (Mm00468865_m1) and Hes6 (Mm00517097_g1). As described previously [ 44 ], the ΔΔCt method was used to quantify the relative abundance of each mRNA. RQ values, calculated only for a threshold cycle (Ct) lower than 37, reflected expression changes in the sample of interest compared with the calibrator sample, after normalization with 18S and Gapdh reference genes.…”

Section: Methodsmentioning

confidence: 99%

“…Then, membranes were blocked using 5% non-fat dried milk (w/v) in TBST (50 mM Tris–HCl, 150 mM NaCl, 0.1% Tween-20, pH 7.4) for 1 h at room temperature, followed by incubation overnight at 4°C with specific primary antibodies diluted in 2.5% non-fat dried milk (w/v) in TBST. The following primary antibodies were used for immunoblotting: 1 : 500 dilution of anti-Pofut1 purified antibody used in a previous study [ 44 ], 1 : 50 dilution of anti-Pax7 antibody (Developmental Studies Hybridoma Bank, University of Iowa, IA), 1 : 2000 dilution of anti-GAPDH (R&D Systems Inc., Minneapolis, MN), 1 : 300 dilution of anti-cleaved NOTCH1 (Val1744; Cell Signaling Technology, Danvers, MA). After three washes in TBST, membranes were incubated for 1 h at room temperature with 1 : 1000 dilution of secondary HRP conjugate antibodies (Dako, Glostrup, Denmark) in 2.5% non-fat dried milk (w/v) in TBST.…”

Section: Methodsmentioning

confidence: 99%

“…In a recent in vitro study, we showed that Pofut1 knockdown reduces Notch signalling and affects differentiation of the mouse myoblast cell line C2C12. The expression patterns of PAX7 and MYOD are modified under these conditions and induce earlier cell differentiation [ 44 ]. In vivo , knock--out of Pofut1 is lethal: mice embryos die at E9.5 with a phenotype similar to that of mice in which NOTCH receptor signalling is inactivated [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

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Reduced Notch signalling leads to postnatal skeletal muscle hypertrophy in Pofut1^cax/caxmice

Jaam¹,

Heu²,

Pennarubia³

et al. 2016

Open Biol.

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Postnatal skeletal muscle growth results from the activation of satellite cells and/or an increase in protein synthesis. The Notch signalling pathway maintains satellite cells in a quiescent state, and once activated, sustains their proliferation and commitment towards differentiation. In mammals, POFUT1-mediated O-fucosylation regulates the interactions between NOTCH receptors and ligands of the DELTA/JAGGED family, thus initiating the activation of canonical Notch signalling. Here, we analysed the consequences of downregulated expression of the Pofut1 gene on postnatal muscle growth in mutant Pofut1cax/cax (cax, compact axial skeleton) mice and differentiation of their satellite cell-derived myoblasts (SCDMs). Pofut1cax/cax mice exhibited muscle hypertrophy, no hyperplasia and a decrease in satellite cell numbers compared with wild-type C3H mice. In agreement with these observations, Pofut1cax/cax SCDMs differentiated earlier concomitant with reduced Pax7 expression and decrease in PAX7+/MYOD− progenitor cells. In vitro binding assays showed a reduced interaction of DELTA-LIKE 1 ligand (DLL1) with NOTCH receptors expressed at the cell surface of SCDMs, leading to a decreased Notch signalling as seen by the quantification of cleaved NICD and Notch target genes. These results demonstrated that POFUT1-mediated O-fucosylation of NOTCH receptors regulates myogenic cell differentiation and affects postnatal muscle growth in mice.

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Section: Discussionsupporting

confidence: 60%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Reduced Notch signalling leads to postnatal skeletal muscle hypertrophy in Pofut1^cax/caxmice

Jaam¹,

Heu²,

Pennarubia³

et al. 2016

Open Biol.

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“…Plagl2 and Pofut1 likely share a promoter region. Pofut1 is an O-fucosylating enzyme that modulates Notch signaling to promote proliferation of muscle stem cells (30). Elucidation of the function of Plagl2 and its relationship with Pofut1 may provide clues to the novel role of Tfr1 in IECs.…”

Section: Discussionmentioning

confidence: 99%

Noncanonical role of transferrin receptor 1 is essential for intestinal homeostasis

Chen

Donovan

Ned-Sykes

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Transferrin receptor 1 (Tfr1) facilitates cellular iron uptake through receptor-mediated endocytosis of iron-loaded transferrin. It is expressed in the intestinal epithelium but not involved in dietary iron absorption. To investigate its role, we inactivated the Tfr1 gene selectively in murine intestinal epithelial cells. The mutant mice had severe disruption of the epithelial barrier and early death. There was impaired proliferation of intestinal epithelial cell progenitors, aberrant lipid handling, increased mRNA expression of stem cell markers, and striking induction of many genes associated with epithelial-to-mesenchymal transition. Administration of parenteral iron did not improve the phenotype. Surprisingly, however, enforced expression of a mutant allele of Tfr1 that is unable to serve as a receptor for iron-loaded transferrin appeared to fully rescue most animals. Our results implicate Tfr1 in homeostatic maintenance of the intestinal epithelium, acting through a role that is independent of its iron-uptake function.transferrin receptor | intestinal epithelium | epithelial-mesenchymal transition | homeostasis | stem cell

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Involvement of ST6Gal I‐mediated α2,6 sialylation in myoblast proliferation and differentiation

et al. 2019

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Myogenesis is a physiological process which involves the proliferation of myoblasts and their differentiation into multinucleated myotubes, which constitute the future muscle fibers. Commitment of myoblasts to differentiation is regulated by the balance between the myogenic factors Pax7 and MyoD. The formation of myotubes requires the presence of glycans, especially N‐glycans, on the cell surface. We examined here the involvement of α2,6 sialylation during murine myoblastic C2C12 cell differentiation by generating a st6gal1‐knockdown C2C12 cell line; these cells exhibit reduced proliferative potential and precocious differentiation due to the low expression of Pax7. The earlier fusion of st6gal1‐knockdown cells leads to a high fusion index and a drop in reserve cells (Pax7+/MyoD−). In st6gal1‐knockdown cells, the Notch pathway is inactivated; consequently, Pax7 expression is virtually abolished, leading to impairment of the proliferation rate. All these results indicate that the decrease in α2,6 sialylation of N‐glycans favors the differentiation of most cells and provokes a significant loss of reserve cells.

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Protein O-Fucosyltransferase 1 Expression Impacts Myogenic C2C12 Cell Commitment via the Notch Signaling Pathway

Cited by 27 publications

References 72 publications

Reduced Notch signalling leads to postnatal skeletal muscle hypertrophy in Pofut1^cax/caxmice

Reduced Notch signalling leads to postnatal skeletal muscle hypertrophy in Pofut1^cax/caxmice

Noncanonical role of transferrin receptor 1 is essential for intestinal homeostasis

Involvement of ST6Gal I‐mediated α2,6 sialylation in myoblast proliferation and differentiation

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Protein O-Fucosyltransferase 1 Expression Impacts Myogenic C2C12 Cell Commitment via the Notch Signaling Pathway

Cited by 27 publications

References 72 publications

Reduced Notch signalling leads to postnatal skeletal muscle hypertrophy in Pofut1cax/caxmice

Reduced Notch signalling leads to postnatal skeletal muscle hypertrophy in Pofut1cax/caxmice

Noncanonical role of transferrin receptor 1 is essential for intestinal homeostasis

Involvement of ST6Gal I‐mediated α2,6 sialylation in myoblast proliferation and differentiation

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Reduced Notch signalling leads to postnatal skeletal muscle hypertrophy in Pofut1^cax/caxmice

Reduced Notch signalling leads to postnatal skeletal muscle hypertrophy in Pofut1^cax/caxmice