USP19-deubiquitinating enzyme regulates levels of major myofibrillar proteins in L6 muscle cells.
The ubiquitin-proteasome system plays an important role in the degradation of myofibrillar proteins that occurs in muscle wasting. Many studies have demonstrated the importance of enzymes mediating conjugation of ubiquitin. However, little is known about the role of deubiquitinating enzymes. We previously showed that the USP19-deubiquitinating enzyme is induced in atrophying skeletal muscle (Combaret L, Adegoke OA, Bedard N, Baracos V, Attaix D, Wing SS. Am J Physiol Endocrinol Metab 288: E693-E700, 2005). To further explore the role of USP19, we used small interfering RNA (siRNA) in L6 muscle cells. Lowering USP19 by 70-90% in myotubes resulted in a 20% decrease in the rate of proteolysis and an 18% decrease in the rate of protein synthesis, with no net change in protein content. Despite the decrease in overall synthesis, there were approximately 1.5-fold increases in protein levels of myosin heavy chain (MHC), actin, and troponin T and a approximately 2.5-fold increase in tropomyosin. USP19 depletion also increased MHC and tropomyosin mRNA levels, suggesting that this effect is due to increased transcription. Consistent with this, USP19 depletion increased myogenin protein and mRNA levels approximately twofold. Lowering myogenin using siRNA prevented the increase in MHC and tropomyosin upon USP19 depletion, indicating that myogenin mediated the increase in myofibrillar proteins. Dexamethasone treatment lowered MHC and increased USP19. Depletion of USP19 reversed the dexamethasone suppression of MHC. These studies demonstrate that USP19 modulates transcription of major myofibrillar proteins and indicate that the ubiquitin system not only mediates the increased protein breakdown but is also involved in the decreased protein synthesis in atrophying skeletal muscle.
Department of Medicine, Polypeptide Laboratory, McGill University and the McGill University Health Centre Research Institute, Montreal, Canada.
Priyanka
P
Sundaram
Zhiyu
Z
Pang
Miao
M
Miao
Lu
L
Yu
Simon S
SS
Wing
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D010450:Endopeptidases / Q000378:metabolism*; D032342:Muscle Cells / Q000201:enzymology / Q000378:metabolism; D009124:Muscle Proteins / Q000096:biosynthesis* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000201:enzymology / Q000378:metabolism*; D009133:Muscular Atrophy / Q000201:enzymology / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D014335:Tropomyosin / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D020107:Troponin T / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
D009124:Muscle Proteins; D034741:RNA, Small Interfering; D014335:Tropomyosin; D020107:Troponin T; D010450:Endopeptidases; C059237:ubiquitin-Nalpha-protein hydrolase; D018995:Myosin Heavy Chains