title: Methods to Monitor and Manipulate TFEB Activity During Autophagy.
abstract: Macroautophagy is a catabolic process deputed to the turnover of intracellular components. Recent studies have revealed that transcriptional regulation is a major mechanism controlling autophagy. Currently, more than 20 transcription factors have been shown to modulate cellular autophagy levels. Among them, the transcription factor EB (TFEB) appears to have the broadest proautophagy role, given its capacity to control the biogenesis of lysosomes and autophagosomes, the two main organelles required for the autophagy pathway. TFEB has attracted major attention owing to its ability to enhance cellular clearance of pathogenic substrates in a variety of animal models of disease, such as lysosomal storage disorders, Parkinson's, Alzheimer's, α1-antitrypsin, obesity as well as others, suggesting that the TFEB pathway represents an extraordinary possibility for future development of innovative therapies. Importantly, the subcellular localization and activity of TFEB are regulated by its phosphorylation status, suggesting that TFEB activity can be pharmacologically targeted. Given the growing list of common and rare diseases in which manipulation of autophagy may be beneficial, in this chapter we describe a set of validated protocols developed to modulate and analyze TFEB-mediated enhancement of autophagy both in vitro and in vivo conditions.
authors.affiliation: Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy.
authors.forename: D L
authors.initials: DL
authors.lastname: Medina
authors.affiliation: Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy; Dulbecco Telethon Institute (DTI), Naples, Italy; Medical Genetics, Federico II University, Naples, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Settembre
authors.affiliation: Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Naples, Italy; Medical Genetics, Federico II University, Naples, Italy; Baylor College of Medicine, Houston, TX, United States; Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, Houston, TX, United States. Electronic address: ballabio@tigem.it.
authors.forename: A
authors.initials: A
authors.lastname: Ballabio
mesh_terms: D000818:Animals; D001343:Autophagy; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D008247:Lysosomes / Q000235:genetics / Q000378:metabolism; D051379:Mice; D061848:Optical Imaging / Q000379:methods; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods; D016707:Tissue Fixation / Q000379:methods; D015533:Transcriptional Activation
chemical_list: D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D012333:RNA, Messenger; C064972:TFEB protein, human
keywords: Autophagy; LSDs; Lysosomal biogenesis; Methods; MiT/TFE; TFE3; TFEB