E3 Ubiquitin Ligase, WWP1, Interacts with AMPKα2 and Down-regulates Its Expression in Skeletal Muscle C2C12 Cells
| Autor: | Sha Jie, Ji Wook Moon, Yong Woo Lee, Nami Kim, Ho Jin Kang, Jung Ok Lee, Sun Hwa Park, Hyeon Soo Kim, Soo Kyung Lee, Ji Hae Kim, Ga Young You, Su Jin Kim, Yongchul Lim |
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| Rok vydání: | 2013 |
| Předmět: |
Male
Snf3 Ubiquitin-Protein Ligases Glucose uptake Down-Regulation Mice Transgenic Biology Biochemistry Mice chemistry.chemical_compound Ubiquitin Cyclic AMP-Dependent Protein Kinase RIIalpha Subunit MG132 medicine Animals Humans Gene Silencing RNA Messenger Muscle Skeletal Protein kinase A Molecular Biology Glutathione Transferase Gene knockdown Skeletal muscle Cell Biology Molecular biology Ubiquitin ligase Mice Inbred C57BL Glucose HEK293 Cells Metabolism medicine.anatomical_structure chemistry Models Animal biology.protein Plasmids |
| Zdroj: | Journal of Biological Chemistry. 288:4673-4680 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m112.406009 |
| Popis: | It is known that the activity of AMP-activated protein kinase (AMPKα2) was depressed under high glucose conditions. However, whether protein expression of AMPKα2 is also down-regulated or not remains unclear. In this study, we showed that the expression of AMPKα2 was down-regulated in cells cultured under high glucose conditions. Treatment of proteasome inhibitor, MG132, blocked high glucose-induced AMPKα2 down-regulation. Endogenous AMPKα2 ubiquitination was detected by immunoprecipitation of AMPKα2 followed by immunoblotting detection of ubiquitin. The yeast-two hybrid (YTH) approach identified WWP1, an E3 ubiquitin ligase, as the AMPKα2-interacting protein in skeletal muscle cells. Interaction between AMPKα2 and WWP1 was validated by co-immunoprecipitation. Knockdown of WWP1 blocked high glucose-induced AMPKα2 down-regulation. The overexpression of WWP1 down-regulated AMPKα2. In addition, the expression of WWP1 is increased under high glucose culture conditions in both mRNA and protein levels. The level of AMPKα2 was down-regulated in the quadriceps muscle of diabetic animal model db/db mice. Expression of WWP1 blocked metformin-induced glucose uptake. Taken together, our results demonstrated that WWP1 down-regulated AMPKα2 under high glucose culture conditions via the ubiquitin-proteasome pathway. |
| Databáze: | OpenAIRE |
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