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

• Published in: Molecular and cellular biology Vol. 35; no. 2; pp. 391 - 405
• Main Authors: Der Vartanian, Audrey, Audfray, Aymeric, Al Jaam, Bilal, Janot, Mathilde, Legardinier, Sébastien, Maftah, Abderrahman, Germot, Agnès
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 01.01.2015; American Society for Microbiology
• Subjects: Animals; Cell Differentiation - physiology; Cell Line; Fucosyltransferases - metabolism; Gene Knockdown Techniques - methods; Life Sciences; Mice; Muscle Development - genetics; Muscle Development - physiology; Myoblasts - cytology; PAX7 Transcription Factor - metabolism; Receptors, Notch - metabolism; Signal Transduction - physiology; MYOBLAST DIFFERENTIATION; LIGAND-BINDING; PAX7; SELF-RENEWAL; FATE DETERMINATION; SATELLITE CELLS; FACTOR-LIKE REPEATS; THROMBOSPONDIN TYPE-1 REPEATS; PRESOMITIC MESODERM; ADULT SKELETAL-MUSCLE
• ISSN: 1098-5549; 0270-7306; 1098-5549
• DOI: 10.1128/MCB.00890-14

The Notch signaling pathway plays a crucial role in skeletal muscle regeneration in mammals by controlling the transition of satellite cells from quiescence to an activated state, their proliferation, and their commitment toward myotubes or self-renewal. O-fucosylation on Notch receptor epidermal growth factor (EGF)-like repeats is catalyzed by the protein O-fucosyltransferase 1 (Pofut1) and primarily controls Notch interaction with its ligands. To approach the role of O-fucosylation in myogenesis, we analyzed a murine myoblastic C2C12 cell line downregulated for Pofut1 expression by short hairpin RNA (shRNA) inhibition during the time course of differentiation. Knockdown of Pofut1 affected the signaling pathway activation by a reduction of the amount of cleaved Notch intracellular domain and a decrease in downstream Notch target gene expression. Depletion in Pax7 + /MyoD − cells and earlier myogenic program entrance were observed, leading to an increase in myotube quantity with a small number of nuclei, reflecting fusion defects. The rescue of Pofut1 expression in knockdown cells restored Notch signaling activation and a normal course in C2C12 differentiation. Our results establish the critical role of Pofut1 on Notch pathway activation during myogenic differentiation.