Nuclear Trapping of the Forkhead Transcription Factor FoxO1 via Sirt-dependent Deacetylation Promotes Expression of Glucogenetic Genes

Activation of NAD-dependent deacetylases, or Sirtuins, prolongs life span and mimics the effects of caloric restriction in yeast. The FoxO subfamily of forkhead transcription factors has been shown to mediate some of the effects of Sirtuins. Here we have shown that Sirtuin activation or hydrogen peroxide treatment overrides the phosphorylation-dependent nuclear exclusion of FoxO1 caused by growth factors and causes nuclear translocation of FoxO1 in hepatocytes. Kinetic measurements of nuclear fluorescence recovery after photobleaching show that FoxO1 is readily diffusible within the nucleus under normal conditions but becomes restricted within a nuclear subdomain following treatment with the prototypical Sirtuin agonist resveratrol or oxidative stress. Expression of FoxO1 target genes is accordingly increased, leading to activation of gluconeogenesis and increased glucose release from hepatocytes. Selective modulation of the FoxO/Sirtuin interaction represents a promising therapeutic modality for metabolic disorders. Activation of NAD-dependent deacetylases, or Sirtuins, prolongs life span and mimics the effects of caloric restriction in yeast. The FoxO subfamily of forkhead transcription factors has been shown to mediate some of the effects of Sirtuins. Here we have shown that Sirtuin activation or hydrogen peroxide treatment overrides the phosphorylation-dependent nuclear exclusion of FoxO1 caused by growth factors and causes nuclear translocation of FoxO1 in hepatocytes. Kinetic measurements of nuclear fluorescence recovery after photobleaching show that FoxO1 is readily diffusible within the nucleus under normal conditions but becomes restricted within a nuclear subdomain following treatment with the prototypical Sirtuin agonist resveratrol or oxidative stress. Expression of FoxO1 target genes is accordingly increased, leading to activation of gluconeogenesis and increased glucose release from hepatocytes. Selective modulation of the FoxO/Sirtuin interaction represents a promising therapeutic modality for metabolic disorders. The long-standing observation that caloric restriction is associated with longevity has led to a widely held theory that metabolism and life span share common cellular pathways (1Wood J.G. Rogina B. Lavu S. Howitz K. Helfand S.L. Tatar M. Sinclair D. Nature. 2004; 430: 686-689Crossref PubMed Scopus (1541) Google Scholar, 2Koubova J. Guarente L. Genes Dev. 2003; 17: 313-321Crossref PubMed Scopus (391) Google Scholar). One such pathway has been proposed to involve forkhead transcription factors of the FoxO subfamily (3Furukawa-Hibi Y. Yoshida-Araki K. Ohta T. Ikeda K. Motoyama N. J. Biol. Chem. 2002; 277: 26729-26732Abstract Full Text Full Text PDF PubMed Scopus (210) Google Scholar, 4van der Horst A. Tertoolen L.G. de Vries-Smits L.M. Frye R.A. Medema R.H. Burgering B.M. J. Biol. Chem. 2004; 279: 28873-28879Abstract Full Text Full Text PDF PubMed Scopus (461) Google Scholar, 5Brunet A. Sweeney L.B. Sturgill J.F. Chua K.F. Greer P.L. Lin Y. Tran H. Ross S.E. Mostoslavsky R. Cohen H.Y. Hu L.S. Cheng H.L. Jedrychowski M.P. Gygi S.P. Sinclair D.A. Alt F.W. Greenberg M.E. Science. 2004; 303: 2011-2015Crossref PubMed Scopus (2570) Google Scholar). Genetic epistasis in Caenorhabditis elegans and metabolic studies in mice indicate that FoxO genes regulate cell differentiation, transformation, and metabolism (6Accili D. Arden K.C. Cell. 2004; 117: 421-426Abstract Full Text Full Text PDF PubMed Scopus (1069) Google Scholar). In C. elegans, mutations of the FoxO ortholog Daf16 rescue the dauer state caused by mutations of the insulin/insulin-like growth factor receptor ortholog Daf2 (7Ogg S. Paradis S. Gottlieb S. Patterson G.I. Lee L. Tissenbaum H.A. Ruvkun G. Nature. 1997; 389: 994-999Crossref PubMed Scopus (1517) Google Scholar, 8Lin K. Dorman J.B. Rodan A. Kenyon C. Science. 1997; 278: 1319-1322Crossref PubMed Scopus (1187) Google Scholar). Moreover, extra copies of the gene encoding the NAD-dependent deacetylase Silent Information Regulator (Sir) 2.1 prolong life span in a Daf16-dependent fashion (9Tissenbaum H.A. Guarente L. Nature. 2001; 410: 227-230Crossref PubMed Scopus (1556) Google Scholar), suggesting that FoxO activity is regulated via deacetylation. These twin observations provide the underpinning for investigations of the role of FoxO proteins in mammalian metabolism and life span. FoxO activity is subject to complex regulation by growth factors and cellular stress. The former inhibit FoxO via serine-threonine phosphorylation and nuclear exclusion (10Kops G.J. Burgering B.M. J. Mol. Med. 1999; 77: 656-665Crossref PubMed Scopus (252) Google Scholar). The latter causes FoxO acetylation, thus promoting the interaction between FoxO and Sirt1, the mammalian ortholog of Sir2.1 (4van der Horst A. Tertoolen L.G. de Vries-Smits L.M. Frye R.A. Medema R.H. Burgering B.M. J. Biol. Chem. 2004; 279: 28873-28879Abstract Full Text Full Text PDF PubMed Scopus (461) Google Scholar, 5Brunet A. Sweeney L.B. Sturgill J.F. Chua K.F. Greer P.L. Lin Y. Tran H. Ross S.E. Mostoslavsky R. Cohen H.Y. Hu L.S. Cheng H.L. Jedrychowski M.P. Gygi S.P. Sinclair D.A. Alt F.W. Greenberg M.E. Science. 2004; 303: 2011-2015Crossref PubMed Scopus (2570) Google Scholar, 11Motta M.C. Divecha N. Lemieux M. Kamel C. Chen D. Gu W. Bultsma Y. McBurney M. Guarente L. Cell. 2004; 116: 551-563Abstract Full Text Full Text PDF PubMed Scopus (1175) Google Scholar, 12Daitoku H. Hatta M. Matsuzaki H. Aratani S. Ohshima T. Miyagishi M. Nakajima T. Fukamizu A. Proc. Natl. Acad. Sci. U. S. A. 2004; 101: 10042-10047Crossref PubMed Scopus (500) Google Scholar). However, the effect of Sirt1-dependent deacetylation on FoxO function remains somewhat controversial, with most (4van der Horst A. Tertoolen L.G. de Vries-Smits L.M. Frye R.A. Medema R.H. Burgering B.M. J. Biol. Chem. 2004; 279: 28873-28879Abstract Full Text Full Text PDF PubMed Scopus (461) Google Scholar, 5Brunet A. Sweeney L.B. Sturgill J.F. Chua K.F. Greer P.L. Lin Y. Tran H. Ross S.E. Mostoslavsky R. Cohen H.Y. Hu L.S. Cheng H.L. Jedrychowski M.P. Gygi S.P. Sinclair D.A. Alt F.W. Greenberg M.E. Science. 2004; 303: 2011-2015Crossref PubMed Scopus (2570) Google Scholar, 12Daitoku H. Hatta M. Matsuzaki H. Aratani S. Ohshima T. Miyagishi M. Nakajima T. Fukamizu A. Proc. Natl. Acad. Sci. U. S. A. 2004; 101: 10042-10047Crossref PubMed Scopus (500) Google Scholar), but not all (11Motta M.C. Divecha N. Lemieux M. Kamel C. Chen D. Gu W. Bultsma Y. McBurney M. Guarente L. Cell. 2004; 116: 551-563Abstract Full Text Full Text PDF PubMed Scopus (1175) Google Scholar), studies suggesting that deacetylation increases FoxO-dependent transcription. In this study, we sought to uncover the mechanism by which stress-induced deacetylation affects FoxO activity. To this end, we studied FoxO translocation using live cell imaging, coupled to measurements of protein kinetics with fluorescence recovery after photobleaching (FRAP) 1The abbreviations used are: FRAP, fluorescence recovery after photobleaching; FLIP, fluorescence loss in photobleaching; GFP, green fluorescent protein; HA, hemagglutinin; PBS, phosphate-buffered saline; NAM, nicotinamide; HDAC, histone deacetylase; TsA, trichostatin A. 1The abbreviations used are: FRAP, fluorescence recovery after photobleaching; FLIP, fluorescence loss in photobleaching; GFP, green fluorescent protein; HA, hemagglutinin; PBS, phosphate-buffered saline; NAM, nicotinamide; HDAC, histone deacetylase; TsA, trichostatin A. and fluorescence loss in photobleaching (FLIP) experiments (13Lippincott-Schwartz J. Altan-Bonnet N. Patterson G.H. Nat. Cell Biol. 2003; : S7-S14PubMed Google Scholar). We also measured expression of FoxO1 target genes and glucose production in hepatocyte cultures. Our findings are consistent with a model in which deacetylation promotes FoxO nuclear retention and increases FoxO-dependent glucose production, thus providing evidence for a mechanism to regulate FoxO activity via subnuclear targeting. DNA Constructs and Cell Transfection—We cloned a mouse FoxO1 cDNA into pEGFP-N1 (Clontech Laboratories) to generate FoxO1-GFP. We used FuGENE 6 (2.5 μg/ml DNA) (Roche Applied Science) to obtain transiently transfected cells expressing FoxO1-GFP. We have described the adenoviral vector expressing HA-tagged wild type FoxO1 in previous publications (14Nakae J. Kitamura T. Silver D.L. Accili D. J. Clin. Investig. 2001; 108: 1359-1367Crossref PubMed Scopus (485) Google Scholar). We transduced cells with adenoviral vectors 24 h before the experiment. Cell Culture—We cultured Sv40 hepatocytes or H4IIE rat hepatoma cells in α-minimal essential medium supplemented with 4% fetal calf serum, 1% Pen-Strep, and 200 nm dexamethasone at 37 °C in a 5% CO2 incubator. We lysed cells in a buffer containing 50 mm Tris-HCl, pH 7.4, 150 mm NaCl, 0.1% SDS, 1% Nonidet P-40, 1% Na deoxycholate, 5 mm Na orthovanadate, 10 mm NaF, 1 mm dithiothreitol, 2 mg/ml pepstatin, 20 μg/ml leupeptin, 10 μg/ml aprotinin, 2 mm phenylmethylsulfonyl fluoride, 10 mm nicotinamide, and 2 μm trichostatin A (TsA). Immunoprecipitation was carried out using anti-HA (12CA5; Roche Applied Science) and anti-GFP monoclonal antibodies (A-11120; Molecular Probes). We processed a 50-μg aliquot of immunoprecipitated proteins for electrophoresis and Western blotting using antisera against FoxO1 (H-128; Santa Cruz), Sirt1 (Upstate Biotechnology), phospho-Ser253 FoxO1, Akt, phospho-Ser473-Akt, or acetyl lysine (Cell Signaling). We employed the following concentrations of reagents: H2O2 0.5 mm, insulin 100 nm, resveratrol 10 μm, nicotinamide 10 mm, and Trichostatin A 2 μm. All were from Sigma. We used Leptomycin B (LC Laboratories) at 20 nm. We cultured HEK 293 cells in minimal essential medium, 10% fetal calf serum, 1% Pen-Strep and transduced them with an adenoviral vector containing HA-tagged wild type FoxO1 24 h before the experiment (14Nakae J. Kitamura T. Silver D.L. Accili D. J. Clin. Investig. 2001; 108: 1359-1367Crossref PubMed Scopus (485) Google Scholar). Image Acquisition and Analysis—We plated and observed cells in LabTek chambers (Nalgene). We captured confocal microscope images on a LSM 510 META microscope (Zeiss) using 488 nm laser excitation for GFP and 568 for rhodamine. Images were captured with a ×63 1.2 water objective and an open pinhole to collect fluorescence from the entire depth of the cell. We analyzed images with the Image J software (NIH). We measured fluorescence separately within the nucleus and the cytoplasm. To calculate relative nuclear fluorescence, we divided nuclear fluorescence by the total amount of cellular fluorescence. Time lapse sequences of images of FoxO1-GFP nuclear translocation were captured with a ×63 1.2 water objective at 10-s intervals (15Lippincott-Schwartz J. Patterson G.H. Science. 2003; 300: 87-91Crossref PubMed Scopus (820) Google Scholar). All from at 20 cells from cells for by FoxO1-GFP by FuGENE 6 (Roche Applied Science) and for Sirt1 in LabTek (Nalgene). We cells in 4% in for using PBS, and them with pH for 10 we cell with and in PBS, and in buffer containing 1% for 10 we cells with Sirt1 (Cell in containing 1% for 1 h at in PBS, and with a for 1 h at were using Analysis—We and observed cells in LabTek chambers (Nalgene). We photobleaching of the nucleus on a LSM 510 META microscope (Zeiss) using 488 nm laser excitation for GFP and a ×63 1.2 water We a in the at laser for we fluorescence recovery by the cell at laser We observed photobleaching we and cells at and were to the used in We recovery from We the total cell fluorescence for using Image J software and the of nuclear fluorescence to the to for the amount the (15Lippincott-Schwartz J. Patterson G.H. Science. 2003; 300: 87-91Crossref PubMed Scopus (820) Google Scholar). All from at 10 cells from Expression using by to We carried out with the to the We carried out using a DNA 2 and were carried out in using or to T. J. Kitamura Y. Y. Arden K.C. Accili D. J. Clin. Investig. 2002; PubMed Scopus Google Scholar). sequences are We employed of from H4IIE glucose production, we the growth medium, cells with and with and α-minimal essential medium for 1 We with and cells in and α-minimal essential medium supplemented with 200 nm 20 μm 2 mm mm and mm Y. M. Accili D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). We measured glucose in using the glucose are the of experiments in FoxO the cellular of FoxO1 using a FoxO1-GFP protein in hepatocytes. FoxO1-GFP to the and in the of serum, in the of the cellular is nuclear and A and These show that the GFP not FoxO of cells with H2O2 to oxidative caused to of FoxO1-GFP to from the to the of the of A and Time that FoxO1-GFP to to the nucleus after in cells and at with a We in cells The in FoxO1 not for by to the the of GFP in the cells was by oxidative The nuclear translocation of FoxO1-GFP peroxide treatment caused by a in activity not for the between of FoxO phosphorylation S. T. Science. 2002; PubMed Scopus Google and nuclear translocation A. Sweeney L.B. Sturgill J.F. Chua K.F. Greer P.L. Lin Y. Tran H. Ross S.E. Mostoslavsky R. Cohen H.Y. Hu L.S. Cheng H.L. Jedrychowski M.P. Gygi S.P. Sinclair D.A. Alt F.W. Greenberg M.E. Science. 2004; 303: 2011-2015Crossref PubMed Scopus (2570) Google in to To this we measured insulin treatment the effect of of insulin FoxO1-GFP to the A and with a not but was to in the of H2O2 A and with we observed that FoxO phosphorylation was in cells FoxO and the mechanism by which FoxO1 to the nucleus oxidative we the effect of that FoxO1 The of NAD-dependent G. Guarente L. 2004; PubMed Scopus Google Scholar), resveratrol Cohen H.Y. Lavu S. J.G. A. B. Sinclair D.A. Nature. 2003; PubMed Scopus Google Scholar), caused FoxO1-GFP translocation to the nucleus A and with a not suggesting that the interaction of FoxO1 with Sirt1 to nuclear experiments that Sirt1 in FoxO1 following H2O2 treatment treatment with the Sirtuin nicotinamide and the histone deacetylase Sirt1 to FoxO1 studies that FoxO1-GFP with Sirt1 in the nucleus H2O2 treatment treatment with resveratrol and H2O2 also caused nuclear translocation A and To the effect of oxidative on FoxO1 is or also we the effects of and on FoxO1-GFP with or to FoxO1-GFP effect on nuclear of FoxO1, the effect of H2O2 A and In treatment FoxO1-GFP nuclear translocation in to H2O2 not Moreover, was to nuclear of treatment not the to resveratrol A and have been shown to FoxO (4van der Horst A. Tertoolen L.G. de Vries-Smits L.M. Frye R.A. Medema R.H. Burgering B.M. J. Biol. Chem. 2004; 279: 28873-28879Abstract Full Text Full Text PDF PubMed Scopus (461) Google Scholar, 5Brunet A. Sweeney L.B. Sturgill J.F. Chua K.F. Greer P.L. Lin Y. Tran H. Ross S.E. Mostoslavsky R. Cohen H.Y. Hu L.S. Cheng H.L. Jedrychowski M.P. Gygi S.P. Sinclair D.A. Alt F.W. Greenberg M.E. Science. 2004; 303: 2011-2015Crossref PubMed Scopus (2570) Google Scholar, 11Motta M.C. Divecha N. Lemieux M. Kamel C. Chen D. Gu W. Bultsma Y. McBurney M. Guarente L. Cell. 2004; 116: 551-563Abstract Full Text Full Text PDF PubMed Scopus (1175) Google Scholar), we sought to the of FoxO1 under with for 2 h increased FoxO1 acetylation, with or resveratrol effect previous FoxO deacetylation by and (4van der Horst A. Tertoolen L.G. de Vries-Smits L.M. Frye R.A. Medema R.H. Burgering B.M. J. Biol. Chem. 2004; 279: 28873-28879Abstract Full Text Full Text PDF PubMed Scopus (461) Google Scholar, 5Brunet A. Sweeney L.B. Sturgill J.F. Chua K.F. Greer P.L. Lin Y. Tran H. Ross S.E. Mostoslavsky R. Cohen H.Y. Hu L.S. Cheng H.L. Jedrychowski M.P. Gygi S.P. Sinclair D.A. Alt F.W. Greenberg M.E. Science. 2004; 303: 2011-2015Crossref PubMed Scopus (2570) Google Scholar, 11Motta M.C. Divecha N. Lemieux M. Kamel C. Chen D. Gu W. Bultsma Y. McBurney M. Guarente L. Cell. 2004; 116: 551-563Abstract Full Text Full Text PDF PubMed Scopus (1175) Google Scholar), findings are consistent with the that are the FoxO in hepatocytes. FoxO1 measured the of FoxO1 using In an of the cell is with a laser and the of from into the is by lapse (13Lippincott-Schwartz J. Altan-Bonnet N. Patterson G.H. Nat. Cell Biol. 2003; : S7-S14PubMed Google Scholar). recovery on the amount of for total cell fluorescence to between cellular following the We the nuclear FoxO1-GFP and the recovery from the In the of serum, nuclear FoxO1-GFP fluorescence within 200 A and after of the nuclear FoxO1-GFP fluorescence nuclear fluorescence We experiments in of photobleaching the nuclear FoxO1-GFP the nucleus the fluorescence A and the of FoxO1-GFP fluorescence These show that FoxO1-GFP is between the and between the nucleus and the of The between the conditions is that the of is in cells 200 phosphorylation-dependent nuclear we measured the of FoxO1-GFP in In the of the nucleus of total cellular FoxO1-GFP fluorescence observed to photobleaching A and of FoxO1-GFP was in the experiments in cells that the nucleus of total fluorescence after A and this to between the nucleus and of FoxO1-GFP to in the The in the of FoxO1-GFP in and cells that FoxO1-GFP is within the nuclear of FoxO1-GFP in we FoxO1 in the of H2O2 or of cells with H2O2 or resveratrol that NAM, but not was to nuclear fluorescence to that FoxO1-GFP is between the nucleus and Sirtuins, but not are a to out from the oxidative on FoxO1-GFP we experiments in cells transfected with GFP and with of photobleaching the nuclear of GFP, the nucleus the fluorescence not These indicate that GFP is between the nucleus and the and that H2O2 not GFP of FoxO using a of FoxO1-GFP fluorescence in the nucleus after experiments in and we experiments (13Lippincott-Schwartz J. Altan-Bonnet N. Patterson G.H. Nat. Cell Biol. 2003; : S7-S14PubMed Google to FoxO1-GFP is to the nuclear photobleaching fluorescence in a of the cell and the fluorescence in the the of a protein readily observed and the of cellular G.H. J. Science. 2002; PubMed Scopus Google Scholar). The of fluorescence loss is on the of the proteins that not between show loss (13Lippincott-Schwartz J. Altan-Bonnet N. Patterson G.H. Nat. Cell Biol. 2003; : S7-S14PubMed Google Scholar). We a of the in cells and captured images between all FoxO1-GFP fluorescence was The not the of nuclear FoxO1-GFP fluorescence, suggesting that a of FoxO1-GFP is to the nuclear and not oxidative stress. experiment was in cells with to a of nuclear FoxO1-GFP fluorescence was after the was We that FoxO1 not between the nucleus and but remains to the nuclear that is a for FoxO1 between the FoxO in or The restriction of FoxO1 to the nuclear oxidative to at FoxO1 kinetics within the nucleus using nuclear In photobleaching a within the nucleus in recovery of fluorescence These indicate that FoxO1-GFP is within the nucleus in In we photobleaching in fluorescence within the to after in cells of a protein or by into a complex or by with nuclear that are such T. Science. 2001; PubMed Scopus Google Scholar). To the kinetics of a complex from a protein to the we in cells with an of nuclear that has been shown to FoxO1 nuclear J. T. Arden K.C. Proc. Natl. Acad. Sci. U. S. A. 1999; PubMed Scopus Google Scholar). in cells an recovery of fluorescence within the These show that FoxO1-GFP is within the nucleus oxidative and resveratrol Sirtuin Activation and of glucose production is an function of FoxO1 (14Nakae J. Kitamura T. Silver D.L. Accili D. J. Clin. Investig. 2001; 108: 1359-1367Crossref PubMed Scopus (485) Google Scholar, J. Kitamura T. Arden K.C. Accili D. Nat. 2002; PubMed Scopus Google Scholar, J. J. Kitamura Y. J. H. Accili D. B.M. Nature. 2003; PubMed Scopus Google Scholar). To the in FoxO1 with we measured glucose production in H4IIE rat hepatoma cells following resveratrol or insulin of cells transduced with wild type FoxO1 in medium supplemented with the and in a in the amount of glucose in the glucose production by resveratrol increased by To the effect of resveratrol was by FoxO1, we transduced cells with nuclear and FoxO1 (14Nakae J. Kitamura T. Silver D.L. Accili D. J. Clin. Investig. 2001; 108: 1359-1367Crossref PubMed Scopus (485) Google Scholar). Expression of the effect of insulin the to resveratrol In glucose release under all conditions We measured of the FoxO1 target genes and growth protein 1 increased expression in cells expressing wild type and but not in cells expressing not in to resveratrol treatment in cells expressing wild type FoxO1 but in cells expressing The and expression expression in to the of These are consistent with a model in which resveratrol increases glucose production by transcription of is that resveratrol increased gene expression in cells expressing a nuclear These findings indicate that nuclear expression not transcription. to an for FoxO1 consistent with a role for deacetylation to transcription. The studies provide evidence for a mechanism by which FoxO1 becomes associated with a nuclear with the nuclear increases transcription and is in increased glucose release from cultured hepatoma The of this is that Sirtuin activation by resveratrol or oxidative FoxO1 within the nuclear and promotes transcription of genes for glucose have been shown to FoxO proteins (4van der Horst A. Tertoolen L.G. de Vries-Smits L.M. Frye R.A. Medema R.H. Burgering B.M. J. Biol. Chem. 2004; 279: 28873-28879Abstract Full Text Full Text PDF PubMed Scopus (461) Google Scholar, 5Brunet A. Sweeney L.B. Sturgill J.F. Chua K.F. Greer P.L. Lin Y. Tran H. Ross S.E. Mostoslavsky R. Cohen H.Y. Hu L.S. Cheng H.L. Jedrychowski M.P. Gygi S.P. Sinclair D.A. Alt F.W. Greenberg M.E. Science. 2004; 303: 2011-2015Crossref PubMed Scopus (2570) Google Scholar, 11Motta M.C. Divecha N. Lemieux M. Kamel C. Chen D. Gu W. Bultsma Y. McBurney M. Guarente L. Cell. 2004; 116: 551-563Abstract Full Text Full Text PDF PubMed Scopus (1175) Google Scholar), we that deacetylation FoxO for nuclear The of oxidative to transcription by a FoxO1 These findings provide evidence that nuclear is but not for activation by FoxO1 and are consistent with the nuclear that FoxO1 is within the nucleus under conditions but becomes restricted in or we J. Accili D. J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google and A. A. Lin Hu L.S. Arden K.C. J. Greenberg M.E. Cell. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar, G.J. de Vries-Smits Burgering B.M. Nature. 1999; PubMed Scopus Google Scholar, S. G. S. Cohen T. J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar, G. S. Cohen J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google have the FoxO We that the of for this The effect of resveratrol in by H2O2 a that is employed to oxidative by concentrations of that the effects of H2O2 in cultured cells are H2O2 is to have effects D. Y. J. Biol. Chem. Full Text PDF PubMed Google Scholar), which increased activity and FoxO1 nuclear exclusion S. T. Science. 2002; PubMed Scopus Google Scholar). is of insulin and FoxO1 becomes nuclear A. Sweeney L.B. Sturgill J.F. Chua K.F. Greer P.L. Lin Y. Tran H. Ross S.E. Mostoslavsky R. Cohen H.Y. Hu L.S. Cheng H.L. Jedrychowski M.P. Gygi S.P. Sinclair D.A. Alt F.W. Greenberg M.E. Science. 2004; 303: 2011-2015Crossref PubMed Scopus (2570) Google Scholar). studies have with the effects of deacetylation on transcription. (4van der Horst A. Tertoolen L.G. de Vries-Smits L.M. Frye R.A. Medema R.H. Burgering B.M. J. Biol. Chem. 2004; 279: 28873-28879Abstract Full Text Full Text PDF PubMed Scopus (461) Google Scholar, 5Brunet A. Sweeney L.B. Sturgill J.F. Chua K.F. Greer P.L. Lin Y. Tran H. Ross S.E. Mostoslavsky R. Cohen H.Y. Hu L.S. Cheng H.L. Jedrychowski M.P. Gygi S.P. Sinclair D.A. Alt F.W. Greenberg M.E. Science. 2004; 303: 2011-2015Crossref PubMed Scopus (2570) Google Scholar, M. H. Hatta M. Matsuzaki H. S. Fukamizu A. J. Mol. Med. 2003; Google Scholar), but not all (11Motta M.C. Divecha N. Lemieux M. Kamel C. Chen D. Gu W. Bultsma Y. McBurney M. Guarente L. Cell. 2004; 116: 551-563Abstract Full Text Full Text PDF PubMed Scopus (1175) Google Scholar), have that deacetylation promotes FoxO-dependent transcription. However, studies on the effects of FoxO proteins on the cell The to the that deacetylation is a of FoxO1 function by providing evidence that regulation of glucose production is also by a of Sirtuins. Our findings are at with of (11Motta M.C. Divecha N. Lemieux M. Kamel C. Chen D. Gu W. Bultsma Y. McBurney M. Guarente L. Cell. 2004; 116: 551-563Abstract Full Text Full Text PDF PubMed Scopus (1175) Google Scholar), increased and expression in mice However, is were in or Moreover, the mice were is that the in and is to Sirt1 to a on this Moreover, that Sirt1 has also been shown to a that is consistent with findings C. W. Gygi S.P. B.M. Nature. PubMed Scopus Google Scholar). is a on Sirtuin modulation are to for the treatment of metabolic disorders. Sirtuin activation increases glucose production in in cultured to the in such cells Sirtuin activation of transcription factors is associated with activity. deacetylation transcription H. Frye R.A. Guarente L. R.A. Cell. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar, S. M. Guarente L. Nature. PubMed Scopus Google Scholar, J. S. Chen D. A. Guarente L. Gu W. Cell. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). in Gu W. Cell Biol. 2003; PubMed Scopus Google Scholar), but is on effect on subnuclear is deacetylation affects FoxO-dependent transcription by The that deacetylation increases the nuclear of FoxO the that deacetylation increases FoxO-dependent but that the of this is to complex FoxO an A. A. Lin Hu L.S. Arden K.C. J. Greenberg M.E. Cell. 1999; Full Text Full Text PDF PubMed Scopus Google and a of gene expression T. J. Kitamura Y. Y. Arden K.C. Accili D. J. Clin. Investig. 2002; PubMed Scopus Google Scholar, J. Kitamura T. Kitamura Y. Arden K.C. Accili D. Dev. Cell. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar). studies with that or to the effects of this on the of FoxO We for and and of the and Accili for We and of the at for