1. The double homeodomain protein DUX4c is associated with regenerating muscle fibers and RNA-binding proteins.
- Author
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Claus, C., Slavin, M., Ansseau, E., Lancelot, C., Bah, K., Lassche, S., Fiévet, M., Greco, A., Tomaiuolo, S., Tassin, A., Dudome, V., Kusters, B., Declèves, A.E., Laoudj-Chenivesse, D., Engelen, B.G.M. van, Nonclercq, D., Belayew, A., Kalisman, N., Coppée, F., Claus, C., Slavin, M., Ansseau, E., Lancelot, C., Bah, K., Lassche, S., Fiévet, M., Greco, A., Tomaiuolo, S., Tassin, A., Dudome, V., Kusters, B., Declèves, A.E., Laoudj-Chenivesse, D., Engelen, B.G.M. van, Nonclercq, D., Belayew, A., Kalisman, N., and Coppée, F.
- Abstract
Item does not contain fulltext, BACKGROUND: We have previously demonstrated that double homeobox 4 centromeric (DUX4C) encoded for a functional DUX4c protein upregulated in dystrophic skeletal muscles. Based on gain- and loss-of-function studies we have proposed DUX4c involvement in muscle regeneration. Here, we provide further evidence for such a role in skeletal muscles from patients affected with facioscapulohumeral muscular dystrophy (FSHD). METHODS: DUX4c was studied at RNA and protein levels in FSHD muscle cell cultures and biopsies. Its protein partners were co-purified and identified by mass spectrometry. Endogenous DUX4c was detected in FSHD muscle sections with either its partners or regeneration markers using co-immunofluorescence or in situ proximity ligation assay. RESULTS: We identified new alternatively spliced DUX4C transcripts and confirmed DUX4c immunodetection in rare FSHD muscle cells in primary culture. DUX4c was detected in nuclei, cytoplasm or at cell-cell contacts between myocytes and interacted sporadically with specific RNA-binding proteins involved, a.o., in muscle differentiation, repair, and mass maintenance. In FSHD muscle sections, DUX4c was found in fibers with unusual shape or central/delocalized nuclei (a regeneration feature) staining for developmental myosin heavy chain, MYOD or presenting intense desmin labeling. Some couples of myocytes/fibers locally exhibited peripheral DUX4c-positive areas that were very close to each other, but in distinct cells. MYOD or intense desmin staining at these locations suggested an imminent muscle cell fusion. We further demonstrated DUX4c interaction with its major protein partner, C1qBP, inside myocytes/myofibers that presented features of regeneration. On adjacent muscle sections, we could unexpectedly detect DUX4 (the FSHD causal protein) and its interaction with C1qBP in fusing myocytes/fibers. CONCLUSIONS: DUX4c upregulation in FSHD muscles suggests it contributes not only to the pathology but also, based on its protein p
- Published
- 2023