O-GlcNAcylation is a key modulator of skeletal muscle sarcomeric morphometry associated to modulation of protein–protein interactions

• Published in: Biochimica et biophysica acta Vol. 1860; no. 9; pp. 2017 - 2030
• Main Authors: Lambert, Matthias, Richard, Elodie, Duban-Deweer, Sophie, Krzewinski, Frederic, Deracinois, Barbara, Dupont, Erwan, Bastide, Bruno, Cieniewski-Bernard, Caroline
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.09.2016; Elsevier
• Series: Biochimica et biophysica acta. General subjects
• Subjects: Actinin - metabolism; Acylation - drug effects; Acylation - physiology; Animals; Cell Line; Crystallins - metabolism; cytoskeleton; Desmin; Desmin - metabolism; Life Sciences; Mice; Microfilament Proteins - metabolism; morphometry; Muscle Contraction - drug effects; Muscle Contraction - physiology; Muscle Fibers, Skeletal - metabolism; Muscle, Skeletal - drug effects; Muscle, Skeletal - metabolism; Myofibrils - metabolism; neuromuscular disorders; O-GlcNAcylation; post-translational modification; Protein Interaction Maps - drug effects; Protein Interaction Maps - physiology; Protein Processing, Post-Translational - drug effects; Protein Processing, Post-Translational - physiology; Protein–protein interactions; proteome; Proteome - metabolism; Pyrans - pharmacology; Sarcomere structure; sarcomeres; Sarcomeres - metabolism; skeletal muscle; Skeletal muscle cells; structural proteins; Thiazoles - pharmacology; αB-crystallin; Sarcomere structure; O-GlcNAcylation; Protein–protein interactions; Skeletal muscle cells; αB-crystallin; Desmin; Mesh:Protein Interaction Maps/physiology; Mesh:Thiazoles/pharmacology; Mesh:Crystallins/metabolism; Mesh:Proteome/metabolism; Mesh:Mice; alpha B-crystallin; Mesh:Microfilament Proteins/metabolism; Mesh:Post-Translational/physiology; Mesh:Desmin/metabolism; Mesh:Protein Processing; Mesh:Actinin/metabolism; Mesh:Myofibrils/metabolism; Mesh:Cell Line; Mesh:Acylation/drug effects; Mesh:Muscle Fibers; Mesh:Muscle Contraction/physiology; Mesh:Pyrans/pharmacology; Mesh:Muscle; Protein-protein interactions; Mesh:Muscle Contraction/drug effects; Mesh:Protein Interaction Maps/drug effects; Mesh:Skeletal/metabolism; Mesh:Skeletal/drug effects; Mesh:Animals; Mesh:Acylation/physiology; Mesh:Post-Translational/drug effects; Mesh:Sarcomeres/metabolism
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2016.06.011

The sarcomere structure of skeletal muscle is determined through multiple protein–protein interactions within an intricate sarcomeric cytoskeleton network. The molecular mechanisms involved in the regulation of this sarcomeric organization, essential to muscle function, remain unclear. O-GlcNAcylation, a post-translational modification modifying several key structural proteins and previously described as a modulator of the contractile activity, was never considered to date in the sarcomeric organization. C2C12 skeletal myotubes were treated with Thiamet-G (OGA inhibitor) in order to increase the global O-GlcNAcylation level. Our data clearly showed a modulation of the O-GlcNAc level more sensitive and dynamic in the myofilament-enriched fraction than total proteome. This fine O-GlcNAc level modulation was closely related to changes of the sarcomeric morphometry. Indeed, the dark-band and M-line widths increased, while the I-band width and the sarcomere length decreased according to the myofilament O-GlcNAc level. Some structural proteins of the sarcomere such as desmin, αB-crystallin, α-actinin, moesin and filamin-C have been identified within modulated protein complexes through O-GlcNAc level variations. Their interactions seemed to be changed, especially for desmin and αB-crystallin. For the first time, our findings clearly demonstrate that O-GlcNAcylation, through dynamic regulations of the structural interactome, could be an important modulator of the sarcomeric structure and may provide new insights in the understanding of molecular mechanisms of neuromuscular diseases characterized by a disorganization of the sarcomeric structure. In the present study, we demonstrated a role of O-GlcNAcylation in the sarcomeric structure modulation. •This paper supports the role of O-GlcNAcylation, a particular glycosylation, in the regulation of interactome, in particular the sarcomeric organization. For the first time, we demonstrated a key role of the O-GlcNAcylation in myofibrillar interactome by the modulation of protein-protein interactions between key structural proteins.•In this paper, e have shown that the interaction between desmin and its molecular chaperone, the αB-crystallin, seems to be modulated according to the global O-GlcNAc level of C2C12 myotubes.•This paper will have interest for glycobiologists (through O-GlcNAcylation), for physiologists and/or cell biologists (through the sarcomeric organization), as well as for proteomists (through interactome).