ABSTRACT
BCL-2 interacting cell death suppressor (BIS), which is ubiquitously expressed, has important roles in various cellular processes, such as apoptosis, the cellular stress response, migration and invasion and protein quality control. In particular, BIS is highly expressed in skeletal and cardiac muscles, and BIS gene mutations result in human myopathy. In this study, we show that mRNA and protein levels of BIS were markedly increased during skeletal myogenesis in C2C12 cells and mouse satellite cells. BIS knockdown did not prevent the early stage of skeletal myogenesis, but did induce muscle atrophy and a decrease in the diameter of myotubes. BIS knockdown significantly suppressed the expression level of myosin heavy chain (MyHC) without changing the expression levels of myogenic marker proteins, such as Mgn, Cav-3 and MG53. In addition, BIS endogenously interacted with MyHC, and BIS knockdown induced MyHC ubiquitination and degradation. From these data, we conclude that molecular association of MyHC and BIS is necessary for MyHC stabilization in skeletal muscle.
Subject(s)
Animals , Humans , Mice , Apoptosis , Cell Death , Muscle Development , Muscle Fibers, Skeletal , Muscle, Skeletal , Muscular Atrophy , Muscular Diseases , Myocardium , Myosin Heavy Chains , Myosins , Quality Control , RNA, Messenger , Ubiquitin , UbiquitinationABSTRACT
BCL-2 interacting cell death suppressor (BIS), which is ubiquitously expressed, has important roles in various cellular processes, such as apoptosis, the cellular stress response, migration and invasion and protein quality control. In particular, BIS is highly expressed in skeletal and cardiac muscles, and BIS gene mutations result in human myopathy. In this study, we show that mRNA and protein levels of BIS were markedly increased during skeletal myogenesis in C2C12 cells and mouse satellite cells. BIS knockdown did not prevent the early stage of skeletal myogenesis, but did induce muscle atrophy and a decrease in the diameter of myotubes. BIS knockdown significantly suppressed the expression level of myosin heavy chain (MyHC) without changing the expression levels of myogenic marker proteins, such as Mgn, Cav-3 and MG53. In addition, BIS endogenously interacted with MyHC, and BIS knockdown induced MyHC ubiquitination and degradation. From these data, we conclude that molecular association of MyHC and BIS is necessary for MyHC stabilization in skeletal muscle.
Subject(s)
Animals , Humans , Mice , Apoptosis , Cell Death , Muscle Development , Muscle Fibers, Skeletal , Muscle, Skeletal , Muscular Atrophy , Muscular Diseases , Myocardium , Myosin Heavy Chains , Myosins , Quality Control , RNA, Messenger , Ubiquitin , UbiquitinationABSTRACT
Extracellular ATP (exATP) has been known to be a critical ligand regulating skeletal muscle differentiation and contractibility. ExATP synthesis was greatly increased with the high level of adenylate kinase 1 (AK1) and ATP synthase beta during C2C12 myogenesis. The exATP synthesis was abolished by the knock-down of AK1 but not by that of ATP synthase beta in C2C12 myotubes, suggesting that AK1 is required for exATP synthesis in myotubes. However, membrane-bound AK1beta was not involved in exATP synthesis because its expression level was decreased during myogenesis in spite of its localization in the lipid rafts that contain various kinds of receptors and mediate cell signal transduction, cell migration, and differentiation. Interestingly, cytoplasmic AK1 was secreted from C2C12 myotubes but not from C2C12 myoblasts. Taken together all these data, we can conclude that AK1 secretion is required for the exATP generation in myotubes.
Subject(s)
Animals , Mice , Adenosine Triphosphate/biosynthesis , Adenylate Kinase/metabolism , Cell Line , Extracellular Space/metabolism , Isoenzymes/metabolism , Muscles/cytologyABSTRACT
Liver, muscle, and adipose tissue are resistant to insulin action in type 2 diabetes. In spite of intensive studies, few diabetic genes have been identified. Recently, mitochondrial impairment has been observed in the muscle and adipose tissues of type 2 diabetes patients, implying that mitochondrial dysfunction could be a pivotal factor in type 2 diabetes. Here, we discuss mitochondrial malfunction leading to type 2 diabetes.
Subject(s)
Humans , Adipose Tissue , Insulin , Liver , MitochondriaABSTRACT
Liver, muscle, and adipose tissue are resistant to insulin action in type 2 diabetes. In spite of intensive studies, few diabetic genes have been identified. Recently, mitochondrial impairment has been observed in the muscle and adipose tissues of type 2 diabetes patients, implying that mitochondrial dysfunction could be a pivotal factor in type 2 diabetes. Here, we discuss mitochondrial malfunction leading to type 2 diabetes.
Subject(s)
Humans , Adipose Tissue , Insulin , Liver , MitochondriaABSTRACT
Mitochondrial biogenesis is known to accompany adipogenesis to complement ATP and acetyl-CoA required for lipogenesis. Here, we demonstrated that mitochondrial proteins such as ATP synthase alpha and beta, and cytochrome c were highly expressed during the 3T3-L1 differentiation into adipocytes. Fully-differentiated adipocytes showed a significant increase of mitochondria under electron microscopy. Analysis by immunofluorescence, cellular fractionation, and surface biotinylation demonstrated the elevated levels of ATP synthase complex found not only in the mitochondria but also on the cell surface (particularly lipid rafts) of adipocytes. High rate of ATP (more than 30 micrometer) synthesis from the added ADP and Pi in the adipocyte media suggests the involvement of the surface ATP synthase complex for the exracellular ATP synthesis. In addition, this ATP synthesis was significantly inhibited in the presence of oligomycin, an ATP synthase inhibitor, and carbonyl cyanide m-chlorophenylhydrazone (CCCP), an ATP synthase uncoupler. Decrease of extracellular ATP synthesis in acidic but not in basic media further indicates that the surface ATP synthase may also be regulated by proton gradient through the plasma membrane.