Abstract
Phosphorylation of FOXO transcription factors is one of the key mechanisms involved in the regulation of the activity, nucleo-cytosolic shuttling, and stability of this family of proteins. Here, we describe several experimental approaches allowing analysis of changes in the phosphorylation of these proteins upon exposure to different stimuli.
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References
Dansen TB, Burgering BM (2008) Unravelling the tumor-suppressive functions of FOXO proteins. Trends Cell Biol 18(9):421–429. https://doi.org/10.1016/j.tcb.2008.07.004
Yang JY, Zong CS, Xia W, Yamaguchi H, Ding Q, Xie X, Lang JY, Lai CC, Chang CJ, Huang WC, Huang H, Kuo HP, Lee DF, Li LY, Lien HC, Cheng X, Chang KJ, Hsiao CD, Tsai FJ, Tsai CH, Sahin AA, Muller WJ, Mills GB, Yu D, Hortobagyi GN, Hung MC (2008) ERK promotes tumorigenesis by inhibiting FOXO3a via MDM2-mediated degradation. Nat Cell Biol 10(2):138–148. https://doi.org/10.1038/ncb1676
Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS, Anderson MJ, Arden KC, Blenis J, Greenberg ME (1999) Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 96(6):857–868
Ho KK, McGuire VA, Koo CY, Muir KW, de Olano N, Maifoshie E, Kelly DJ, McGovern UB, Monteiro LJ, Gomes AR, Nebreda AR, Campbell DG, Arthur JS, Lam EW (2012) Phosphorylation of FOXO3a on Ser-7 by p38 promotes its nuclear localization in response to doxorubicin. J Biol Chem 287(2):1545–1555. https://doi.org/10.1074/jbc.M111.284224
Greer EL, Dowlatshahi D, Banko MR, Villen J, Hoang K, Blanchard D, Gygi SP, Brunet A (2007) An AMPK-FOXO pathway mediates longevity induced by a novel method of dietary restriction in C. elegans. Curr Biol 17(19):1646–1656. https://doi.org/10.1016/j.cub.2007.08.047
Lehtinen MK, Yuan Z, Boag PR, Yang Y, Villen J, Becker EB, DiBacco S, de la Iglesia N, Gygi S, Blackwell TK, Bonni A (2006) A conserved MST-FOXO signaling pathway mediates oxidative-stress responses and extends life span. Cell 125(5):987–1001. https://doi.org/10.1016/j.cell.2006.03.046
Davila D, Connolly NM, Bonner H, Weisova P, Dussmann H, Concannon CG, Huber HJ, Prehn JH (2012) Two-step activation of FOXO3 by AMPK generates a coherent feed-forward loop determining excitotoxic cell fate. Cell Death Differ 19(10):1677–1688. https://doi.org/10.1038/cdd.2012.49
Salazar M, Lorente M, Garcia-Taboada E, Perez Gomez E, Davila D, Zuniga-Garcia P, Maria Flores J, Rodriguez A, Hegedus Z, Mosen-Ansorena D, Aransay AM, Hernandez-Tiedra S, Lopez-Valero I, Quintanilla M, Sanchez C, Iovanna JL, Dusetti N, Guzman M, Francis SE, Carracedo A, Kiss-Toth E, Velasco G (2015) Loss of Tribbles pseudokinase-3 promotes Akt-driven tumorigenesis via FOXO inactivation. Cell Death Differ 22(1):131–144. https://doi.org/10.1038/cdd.2014.133
Essafi A, Gomes AR, Pomeranz KM, Zwolinska AK, Varshochi R, McGovern UB, Lam EW (2009) Studying the subcellular localization and DNA-binding activity of FoxO transcription factors, downstream effectors of PI3K/Akt. Methods Mol Biol 462:201–211. https://doi.org/10.1007/978-1-60327-115-8_13
Acknowledgments
Work in G Velasco’s group is funded by the PI15/00339 grant, integrated into the State Plan for R & D + I2013-2016 and funded by the Instituto de salud Carlos III (ISCIII) and the European Regional Development Fund (ERDF), by the GA 721532 Marie Sklowdoska-Curie Innovative Training Network (ITN) action TRAIN (Tribbles research and Innovation Network) funded by the H2020 program of the European Commission and by a grant from Voices Against Brain Cancer. We than all member of the L1-L2 groups for their contribution to the setup of the experimental approaches described in this chapter.
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Orea-Soufi, A., Dávila, D., Salazar-Roa, M., de Mar Lorente, M., Velasco, G. (2019). Phosphorylation of FOXO Proteins as a Key Mechanism to Regulate Their Activity. In: Link, W. (eds) FOXO Transcription Factors. Methods in Molecular Biology, vol 1890. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8900-3_5
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DOI: https://doi.org/10.1007/978-1-4939-8900-3_5
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