title: SIRT1-SIRT3 Axis Regulates Cellular Response to Oxidative Stress and Etoposide.
abstract: Sirtuins are conserved NAD+-dependent deacylases. SIRT1 is a nuclear and cytoplasmic sirtuin involved in the control of histones a transcription factors function. SIRT3 is a mitochondrial protein, which regulates mitochondrial function. Although, both SIRT1 and SIRT3 have been implicated in resistance to cellular stress, the link between these two sirtuins has not been studied so far. Here we aimed to unravel: i) the role of SIRT1-SIRT3 axis for cellular response to oxidative stress and DNA damage; ii) how mammalian cells modulate such SIRT1-SIRT3 axis and which mechanisms are involved. Therefore, we analyzed the response to different stress stimuli in WT or SIRT1-silenced cell lines. Our results demonstrate that SIRT1-silenced cells are more resistant to H2O2and etoposide treatment showing decreased ROS accumulation, γ-H2AX phosphorylation, caspase-3 activation and PARP cleavage. Interestingly, we observed that SIRT1-silenced cells show an increased SIRT3 expression. To explore such a connection, we carried out luciferase assays on SIRT3 promoter demonstrating that SIRT1-silencing increases SIRT3 promoter activity and that such an effect depends on the presence of SP1 and ZF5 recognition sequences on SIRT3 promoter. Afterwards, we performed co-immunoprecipitation assays demonstrating that SIRT1 binds and deacetylates the transcription inhibitor ZF5 and that there is a decreased interaction between SP1 and ZF5 in SIRT1-silenced cells. Therefore, we speculate that acetylated ZF5 cannot bind and sequester SP1 that is free, then, to increase SIRT3 transcription. In conclusion, we demonstrate that cells with low SIRT1 levels can maintain their resistance and survival by increasing SIRT3 expression. J. Cell. Physiol. 232: 1835-1844, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Experimental Medicine, University of Rome, Sapienza, Rome, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Carnevale
authors.affiliation: Department of Neurorehabilitation Sciences, Casa Cura Policlinico (CCP), Milan, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Pellegrini
authors.affiliation: Department of Biology, Ecology and Earth Sciences, University of Calabria, Rende, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: D'Aquila
authors.affiliation: Department of Experimental Medicine, University of Rome, Sapienza, Rome, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Saladini
authors.affiliation: Department of Experimental Medicine, University of Rome, Sapienza, Rome, Italy.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Lococo
authors.affiliation: Department of Experimental Medicine, University of Rome, Sapienza, Rome, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Polletta
authors.affiliation: The Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, Nebraska.
authors.forename: Enza
authors.initials: E
authors.lastname: Vernucci
authors.affiliation: Department of Experimental Medicine, University of Rome, Sapienza, Rome, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Foglio
authors.affiliation: Physics of Life Processes, Kammerlingh Onnes-Huygens Laboratory, Leiden University, Leiden, The Netherlands.
authors.forename: Stefano
authors.initials: S
authors.lastname: Coppola
authors.affiliation: Department of Cellular and Molecular Pathology, IRCCS San Raffaele, Rome, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Sansone
authors.affiliation: Department of Biology, Ecology and Earth Sciences, University of Calabria, Rende, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Passarino
authors.affiliation: Department of Biology, Ecology and Earth Sciences, University of Calabria, Rende, Italy.
authors.forename: Dina
authors.initials: D
authors.lastname: Bellizzi
authors.affiliation: Consortium MEBIC, San Raffaele University, Rome, Italy.
authors.forename: Matteo A
authors.initials: MA
authors.lastname: Russo
authors.affiliation: Department of Cellular and Molecular Pathology, IRCCS San Raffaele, Rome, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Fini
authors.affiliation: Department of Experimental Medicine, University of Rome, Sapienza, Rome, Italy.|Department of Cellular and Molecular Pathology, IRCCS San Raffaele, Rome, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Tafani
mesh_terms: D000107:Acetylation / Q000187:drug effects; D000818:Animals; D045744:Cell Line, Tumor; D019610:Cytoprotection / Q000187:drug effects; D005047:Etoposide / Q000494:pharmacology*; D020868:Gene Silencing / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D018384:Oxidative Stress / Q000187:drug effects*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D056564:Sirtuin 1 / Q000378:metabolism*; D056566:Sirtuin 3 / Q000378:metabolism*; D016329:Sp1 Transcription Factor / Q000378:metabolism
chemical_list: D017382:Reactive Oxygen Species; D016329:Sp1 Transcription Factor; D005047:Etoposide; D056564:Sirtuin 1; D056566:Sirtuin 3