1 Scientific RepoRts | 7:45076 | DOI: 10.1038/srep45076 www.nature.com/scientificreports Acute and chronic mitochondrial respiratory chain defciency diferentially regulate lysosomal biogenesis Lorena Fernández-Mosquera 1,2 , Cátia V. Diogo 1 , King Faisal Yambire 1,3 , Gabriela L. santos 1 , Marta Luna sánchez 4 , paule Bénit 5,6 , pierre Rustin 5,6 , Luis Carlos Lopez 4 , Ira Milosevic 7 & Nuno Raimundo 1 Mitochondria are key cellular signaling platforms, afecting fundamental processes such as cell proliferation, diferentiation and death. However, it remains unclear how mitochondrial signaling afects other organelles, particularly lysosomes. Here, we demonstrate that mitochondrial respiratory chain (RC) impairments elicit a stress signaling pathway that regulates lysosomal biogenesis via the microphtalmia transcription factor family. Interestingly, the efect of mitochondrial stress over lysosomal biogenesis depends on the timeframe of the stress elicited: while RC inhibition with rotenone or uncoupling with CCCP initially triggers lysosomal biogenesis, the efect peaks after few hours and returns to baseline. Long-term RC inhibition by long-term treatment with rotenone, or patient mutations in fbroblasts and in a mouse model result in repression of lysosomal biogenesis. The induction of lysosomal biogenesis by short-term mitochondrial stress is dependent on TFEB and MITF, requires AMPK signaling and is independent of calcineurin signaling. These results reveal an integrated view of how mitochondrial signaling afects lysosomes, which is essential to fully comprehend the consequences of mitochondrial malfunction, particularly in the context of mitochondrial diseases. Mitochondria are fundamental organelles in metabolism and cell biology 1 . In addition to the long characterized metabolic roles, including energy metabolism and citrate cycle, iron and calcium metabolism and regulation of apoptosis, mitochondria are increasingly recognized as key signaling platforms afecting not only major cellular processes but also far-reaching systemic regulatory mechanisms 2 . Furthermore, it is now clear that mitochon- dria interact with other organelles, both through physical contact sites and through signaling pathways 3,4 . Te interaction between mitochondria and the endoplasmic reticulum (ER) provides pivotal examples of both: the mitochondrial-ER contact sites have been extensively studied and are known to be key players in cellular calcium homeostasis 5 , while mitochondrial dysfunction has also been shown to trigger ER stress 6 . Recently, it was also reported that acute mitochondrial malfunction results in impairment of lysosomes and induction of lysosomal biogenesis 7 . Te lysosomes are part of the endolysosomal system, and have traditionally been associated with degradation of cellular components 8,9 . In the recent years, the lysosomes have been shown to be much more than an acid recycling bag, as their role in amino acid sensing and autophagy regulation has been elucidated 8,9 . Furthermore, a lysosomal stress response that increases lysosomal biogenesis was also identifed 10,11 . Tis stress response is medi- ated by the transcription factors of the microphtalmia family, TFEB, MITF, TFEC and TFE3 10–12 . Tese transcrip- tion factors are associated to the lysosome during basal conditions, where they become phosphorylated by the mTORC1 complex. Upon lysosomal stress or amino acid starvation, mTORC1 is inactivated and Ca 2+ is released 1 institute of cellular Biology, University Medical center Goettingen, Goettingen, Germany. 2 Doctoral Program on Molecular Medicine, University of Goettingen, Goettingen, Germany. 3 international Max-Planck Research School on neuroscience, Goettingen, Germany. 4 Departamento de fisiología, facultad de Medicina and instituto de Biotecnología, centro de investigación Biomédica, University of Granada, Granada, Spain. 5 INSERM UMR 1141, Hôpital Robert Debré, Paris, france. 6 Faculté de Médecine Denis Diderot, Université Paris Diderot – Paris 7, Site Robert Debré, Paris, france. 7 european neuroscience institute, Goettingen, Germany. correspondence and requests for materials should be addressed to n.R. (email: nuno.raimundo@med.uni-goettingen.de) Received: 27 October 2016 accepted: 17 February 2017 Published: 27 March 2017 OPEN OPEN