Transcriptional and post-translational activation of AMPKα by oxidative, heat, and cold stresses in the red flour beetle, Tribolium castaneum
The AMP-activated protein kinase (AMPK) has important roles in the regulation of energy metabolism, and AMPK activity and its regulation have been the focus of relevant investigations. However, functional characterization of AMPK is still limited in insects. In this study, the full-length cDNA coding AMPKα (TcAMPKα) was isolated from the red flour beetle, Tribolium castaneum. The TcAMPKα gene contains an ORF of 1581 bp encoding a protein of 526 amino acid residues, which shared conserved domain structure with Drosophila melanogaster and mammalian orthologs. Exposure of female adults to oxidative, heat, and cold stresses caused an increase in TcAMPKα mRNA expression levels and phosphorylation of Thr-173 in the activation loop. The RNAi-mediated knockdown of TcAMPKα resulted in the increased sensitivity of T. castaneum to oxidative, heat, and cold stresses. These results suggest that stress signals regulate TcAMPKα activity, and TcAMPKα plays an important role in enabling protective mechanisms and processes that confer resistance to environmental stress.
KeywordsTribolium castaneum AMPK Stress resistance Phosphorylation RNAi
JW conceived and designed research. HJ, NZ, MC, CJ, HG, and XX conducted experiments. XM, FD, LM, XY, and KQ analyzed data. HJ and JW wrote the manuscript. All authors read and approved the manuscript.
This work was supported by the National Natural Science Foundation of China under grant nos. 31572000 and 31871974.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Chen L, Xin FJ, Wang J, Hu J, Zhang YY, Wan S, Cao LS, Lu C, Li P, Yan SF, Neumann D, Schlattner U, Xia B, Wang ZX, Wu JW (2013) Conserved regulatory elements in AMPK. Nature 498:E8–E10. https://doi.org/10.1038/nature12189
- Crozet P, Margalha L, Confraria A, Rodrigues A, Martinho C, Adamo M, Elias CA, Baena-González E (2014) Mechanisms of regulation of SNF1/AMPK/SnRK1 protein kinases. Front Plant Sci 5(190). https://doi.org/10.3389/fpls.2014.00190
- Hardie DG, Carling D, Halford N (1994) Roles of the Snf1/Rkin1/AMP-activated protein kinase family in the response to environmental and nutritional stress. Seminars in Cell Biology 5(6):409–416. https://doi.org/10.1006/scel.1994.1048
- Hawley SA, Davison M, Woods A, Davies SP, Beri RK, Carling D, Hardie DG (1996) Characterization of the amp-activated protein kinase kinase from rat liver and identification of threonine 172 as the major site at which it phosphorylates amp-activated protein kinase. J Biol Chem 271:27879–27887. https://doi.org/10.1074/jbc.271.44.27879 CrossRefGoogle Scholar
- He X, Lu Z, Ma B, Zhang L, Li J, Jiang Y, Zhou G, Gao F (2018) Chronic heat stress damages small intestinal epithelium cells associated with the adenosine 5′-monophosphate-activated protein kinase pathway in broilers. J Agric Food Chem 66:7301–7309. https://doi.org/10.1021/acs.jafc.8b02145 CrossRefGoogle Scholar
- Kosztelnik M, Kurucz A, Papp D, Jones E, Sigmond T, Barna J, Traka MH, Lorincz T, Szarka A, Banhegyi G, Vellai T, Korcsmaros T, Kapuy O (2019) Suppression of AMPK/aak-2 by NRF2/SKN-1 down-regulates autophagy during prolonged oxidative stress. FASEB J 33:2372–2387. https://doi.org/10.1096/fj.201800565RR CrossRefGoogle Scholar
- She C, Zhu LQ, Zhen YF, Wang XD, Dong QR (2014) Activation of AMPK protects against hydrogen peroxide-induced osteoblast apoptosis through autophagy induction and NADPH maintenance: new implications for osteonecrosis treatment? Cell Signal 26:1–8. https://doi.org/10.1016/j.cellsig.2013.08.046 CrossRefGoogle Scholar
- Suzuki A, Okamoto S, Lee S, Saito K, Shiuchi T, Minokoshi Y (2007) Leptin stimulates fatty acid oxidation and peroxisome proliferator-activated receptor α gene expression in mouse C2C12 myoblasts by changing the subcellular localization of the α2 form of AMP-activated protein kinase. Mol Cell Biol 27:4317–4327. https://doi.org/10.1128/MCB.02222-06 CrossRefGoogle Scholar
- Thompson-Jaeger S, Francois J, Gaughran JP, Tatchell K (1991) Deletion of SNF1 affects the nutrient response of yeast and resembles mutations which activate the adenylate cyclase pathway. Genetics. 129:697–706Google Scholar
- Wilson WA, Hawley SA, Hardie DG (1996) Glucose repression/derepression in budding yeast: SNF1 protein kinase is activated by phosphorylation under derepressing conditions, and this correlates with a high AMP: ATP ratio. Curr Biol 6:1426–1434. https://doi.org/10.1016/S0960-9822(96)00747-6 CrossRefGoogle Scholar