title: Regulation of SIRT2-dependent α-tubulin deacetylation by cellular NAD levels.
abstract: Acetylation of α-tubulin on lysine 40 is one of the major posttranslational modifications of microtubules. The acetylation reaction is catalyzed by alpha-tubulin N-acetyltransferase and the modification can be reversed by either the NAD-independent class II histone deacetylase HDAC6 or the NAD-dependent deacetylase SIRT2. In this study, we assessed to what extent cellular NAD levels are involved in the regulation of the α-tubulin acetylation state. Cells were subjected to different treatments known to influence cellular NAD content. In response to NAD depletion caused by inhibition of NAD synthesis from nicotinamide, α-tubulin was hyperacetylated. Under these conditions, the normal tubulin acetylation state could be restored by providing the cells with alternative NAD precursors. Likewise, decreasing the rate of endogenous NAD consumption using an inhibitor of poly-ADP-ribosylation also stabilized the acetylation of α-tubulin. Conversely, the level of acetylated α-tubulin decreased when NAD synthesis was enhanced by overexpression of an NAD biosynthetic enzyme. Combined, these results show that the tubulin acetylation status is reciprocally regulated by cellular NAD levels. Furthermore, we provide evidence confirming that the NAD-dependent regulation of tubulin acetylation is mediated by SIRT2.
authors.affiliation: Department of Molecular Biology, University of Bergen, Postbox 7803, 5020 Bergen, Norway.
authors.forename: Renate Hvidsten
authors.initials: RH
authors.lastname: Skoge
authors.affiliation: Department of Molecular Biology, University of Bergen, Postbox 7803, 5020 Bergen, Norway.
authors.forename: Christian
authors.initials: C
authors.lastname: Dölle
authors.affiliation: Department of Molecular Biology, University of Bergen, Postbox 7803, 5020 Bergen, Norway. Electronic address: mathias.ziegler@mbi.uib.no.
authors.forename: Mathias
authors.initials: M
authors.lastname: Ziegler
mesh_terms: D000107:Acetylation; D000178:Acrylamides / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D057809:HEK293 Cells / Q000187:drug effects / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008239:Lysine / Q000378:metabolism; D009243:NAD / Q000096:biosynthesis / Q000378:metabolism*; D010616:Phenanthrenes / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology; D000071137:Poly (ADP-Ribose) Polymerase-1; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D056565:Sirtuin 2 / Q000235:genetics / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism*
chemical_list: D000178:Acrylamides; D004791:Enzyme Inhibitors; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; C480543:N-(4-(1-benzoylpiperidin-4-yl)butyl)-3-(pyridin-3-yl)acrylamide; C434926:N-(oxo-5,6-dihydrophenanthridin-2-yl)-N,N-dimethylacetamide hydrochloride; D010616:Phenanthrenes; D010880:Piperidines; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D014404:Tubulin; D009243:NAD; C012589:trichostatin A; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; C474199:SIRT2 protein, human; D056565:Sirtuin 2; D008239:Lysine
keywords: NAD metabolism; NAD-dependent deacetylation; Sirtuin; Tubulin acetylation