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Ca2+-Dependent Regulation of TFEB and Lysosomal Function

  • Simone Di Paola
  • Diego L. MedinaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1925)

Abstract

Lysosomes are emerging as calcium store organelles that can modulate various intracellular processes such as the regulation of nutrient signaling through the activation of TFEB, a master gene for lysosomal function, or very specialized functions like lysosomal exocytosis. Here, we describe two different techniques that can be used to study these processes. In the case report, we described two studies where these methodologies allowed us to unravel the role of calcineurin in the dephosphorylation of TFEB as well as the involvement of TFEB in lysosomal exocytosis, respectively.

Key words

Lysosomes Ca2 + TFEB Calcineurin Lysosomal exocytosis 

Notes

Acknowledgments

This work was supported by grants from the ML4 Foundation (SDP and DLM) and Cure Sanfilippo Foundation (DLM) and by EU Horizon 2020, BATCure 666918 (DLM).

References

  1. 1.
    Settembre C, Fraldi A, Medina DL, Ballabio A (2013) Signals from the lysosome: a control centre for cellular clearance and energy metabolism. Nat Rev Mol Cell Biol 14(5):283–296.  https://doi.org/10.1038/nrm3565 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Sardiello M, Palmieri M, di Ronza A, Medina DL, Valenza M, Gennarino VA, Di Malta C, Donaudy F, Embrione V, Polishchuk RS, Banfi S, Parenti G, Cattaneo E, Ballabio A (2009) A gene network regulating lysosomal biogenesis and function. Science 325(5939):473–477CrossRefGoogle Scholar
  3. 3.
    Medina DL, Fraldi A, Bouche V, Annunziata F, Mansueto G, Spampanato C, Puri C, Pignata A, Martina JA, Sardiello M, Palmieri M, Polishchuk R, Puertollano R, Ballabio A (2011) Transcriptional activation of lysosomal exocytosis promotes cellular clearance. Dev Cell 21(3):421–430CrossRefGoogle Scholar
  4. 4.
    Settembre C, Di Malta C, Polito VA, Garcia Arencibia M, Vetrini F, Erdin S, Erdin SU, Huynh T, Medina D, Colella P, Sardiello M, Rubinsztein DC, Ballabio A (2011) TFEB links autophagy to lysosomal biogenesis. Science 332(6036):1429–1433CrossRefGoogle Scholar
  5. 5.
    Settembre C, Zoncu R, Medina DL, Vetrini F, Erdin S, Erdin S, Huynh T, Ferron M, Karsenty G, Vellard MC, Facchinetti V, Sabatini DM, Ballabio A (2012) A lysosome-to-nucleus signalling mechanism senses and regulates the lysosome via mTOR and TFEB. EMBO J 31(5):1095–1108CrossRefGoogle Scholar
  6. 6.
    Ferron M, Settembre C, Shimazu J, Lacombe J, Kato S, Rawlings DJ, Ballabio A, Karsenty G (2013) A RANKL-PKCbeta-TFEB signaling cascade is necessary for lysosomal biogenesis in osteoclasts. Genes Dev 27(8):955–969CrossRefGoogle Scholar
  7. 7.
    Medina DL, Di Paola S, Peluso I, Armani A, De Stefani D, Venditti R, Montefusco S, Scotto-Rosato A, Prezioso C, Forrester A, Settembre C, Wang W, Gao Q, Xu H, Sandri M, Rizzuto R, De Matteis MA, Ballabio A (2015) Lysosomal calcium signalling regulates autophagy through calcineurin and TFEB. Nat Cell Biol 17(3):288–299CrossRefGoogle Scholar
  8. 8.
    Spampanato C, Feeney E, Li L, Cardone M, Lim JA, Annunziata F, Zare H, Polishchuk R, Puertollano R, Parenti G, Ballabio A, Raben N (2013) Transcription factor EB (TFEB) is a new therapeutic target for Pompe disease. EMBO Mol Med 5(5):691–706CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Telethon Institute of Genetics and Medicine (TIGEM)PozzuoliItaly

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