Abstract
Ubiquitin-mediated protein degradation is the main mechanism for controlled proteolysis, which is crucial for muscle development and maintenance. The ankyrin repeat-containing protein with a suppressor of cytokine signaling box 2 gene (ASB2) encodes the specificity subunit of an E3 ubiquitin ligase complex involved in differentiation of hematopoietic cells. Here, we provide the first evidence that a novel ASB2 isoform, ASB2β, is important for muscle differentiation. ASB2β is expressed in muscle cells during embryogenesis and in adult tissues. ASB2β is part of an active E3 ubiquitin ligase complex and targets the actin-binding protein filamin B (FLNb) for proteasomal degradation. Thus, ASB2β regulates FLNb functions by controlling its degradation. Knockdown of endogenous ASB2β by shRNAs during induced differentiation of C2C12 cells delayed FLNb degradation as well as myoblast fusion and expression of muscle contractile proteins. Finally, knockdown of FLNb in ASB2β knockdown cells restores myogenic differentiation. Altogether, our results suggest that ASB2β is involved in muscle differentiation through the targeting of FLNb to destruction by the proteasome.
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Accession codes
Abbreviations
- ASB2 :
-
ankyrin repeat-containing protein with a suppressor of cytokine signaling box 2 gene
- CRL:
-
Cullin RING ligase
- FLN:
-
filamin
- HECT:
-
homologous to the E6-associated protein C terminus
- MHC:
-
myosin heavy chain
- MURF:
-
muscle RING finger
- RING:
-
really interesting new gene
- SOCS:
-
suppressor of cytokine signaling
- UIM:
-
ubiquitin-interacting motif
- UPS:
-
ubiquitin–proteasome system
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Acknowledgements
We thank J Dubrulle and O Pourquié for the chicken ASB2 probe and initial help with in situ hybridizations, M-A Bonnin for technical assistance with in situ hybridizations and V Mouly and the human cell culture platform from the Myology Institute in Paris for providing human primary myoblasts. We are grateful to D Heard for the design of shRNAs directed against mouse ASB2β. We thank Lucie Carrier for critical reading of the article. This work was supported by the Centre National de la Recherche Scientifique (CNRS), the Université de Toulouse, the Université Pierre et Marie Curie and by grants to DAC from the National Institutes of Health (GM068600 and HL089433), to CML from the Université Paul Sabatier and to PGL from the Agence Nationale de la Recherche (Programme Jeunes Chercheuses, Jeunes Chercheurs), the Association Française contre les Myopathies, the Association pour la Recherche sur le Cancer (Programme Equipe Nouvelle) and the Fondation pour la Recherche Médicale (Programme Installation d'une nouvelle équipe). NF Bello is supported by the Association Française contre les Myopathies. ML Heuzé was supported by a doctoral Allocation de Recherche du Ministère de la Recherche et des Technologies and by the Association pour la Recherche sur le Cancer.
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Bello, N., Lamsoul, I., Heuzé, M. et al. The E3 ubiquitin ligase specificity subunit ASB2β is a novel regulator of muscle differentiation that targets filamin B to proteasomal degradation. Cell Death Differ 16, 921–932 (2009). https://doi.org/10.1038/cdd.2009.27
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DOI: https://doi.org/10.1038/cdd.2009.27
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