title: Phosphorylation regulates SIRT1 function.
abstract: BACKGROUND
  SIR2 is an NAD(+)-dependent deacetylase [1]-[3] implicated in the regulation of lifespan in species as diverse as yeast [4], worms [5], and flies [6]. We previously reported that the level of SIRT1, the mammalian homologue of SIR2 [7], [8], is coupled to the level of mitotic activity in cells both in vitro and in vivo[9]. Cells from long-lived mice maintained SIRT1 levels of young mice in tissues that undergo continuous cell replacement by proliferating stem cells. Changes in SIRT1 protein level were not associated with changes in mRNA level, suggesting that SIRT1 could be regulated post-transcriptionally. However, other than a recent report on sumoylation [10] and identification of SIRT1 as a nuclear phospho-protein by mass spectrometry [11], post-translational modifications of this important protein have not been reported.
  
  
  RESULTS
  We identified 13 residues in SIRT1 that are phosphorylated in vivo using mass spectrometry. Dephosphorylation by phosphatases in vitro resulted in decreased NAD(+)-dependent deacetylase activity. We identified cyclinB/Cdk1 as a cell cycle-dependent kinase that forms a complex with and phosphorylates SIRT1. Mutation of two residues phosphorylated by Cyclin B/Cdk1 (threonine 530 and serine 540) disturbs normal cell cycle progression and fails to rescue proliferation defects in SIRT1-deficient cells [12], [13].
  
  
  CONCLUSIONS
  Pharmacological manipulation of SIRT1 activity is currently being tested as a means of extending lifespan in mammals. Treatment of obese mice with resveratrol, a pharmacological activator of SIRT1, modestly but significantly improved longevity and, perhaps more importantly, offered some protection against the development of type 2 diabetes mellitus and metabolic syndrome [14]-[16]. Understanding the endogenous mechanisms that regulate the level and activity of SIRT1, therefore, has obvious relevance to human health and disease. Our results identify phosphorylation by cell cycle dependent kinases as a major mechanism controlling the level and function of this sirtuin and complement recent reports of factors that inhibit [17], [18] and activate [19] SIRT1 by protein-protein interactions.
authors.affiliation: Neuroscience Graduate Program, University of Virginia, Charlottesville, Virginia, United States of America.
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Sasaki
authors.forename: Bernhard
authors.initials: B
authors.lastname: Maier
authors.forename: Katarzyna D
authors.initials: KD
authors.lastname: Koclega
authors.forename: Maksymilian
authors.initials: M
authors.lastname: Chruszcz
authors.forename: Wendy
authors.initials: W
authors.lastname: Gluba
authors.forename: P Todd
authors.initials: PT
authors.lastname: Stukenberg
authors.forename: Wladek
authors.initials: W
authors.lastname: Minor
authors.forename: Heidi
authors.initials: H
authors.lastname: Scrable
mesh_terms: D020816:Amino Acid Motifs / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism; D020134:Catalytic Domain; D002478:Cells, Cultured; D019926:Cyclin B / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D016213:Cyclins / Q000378:metabolism; D004789:Enzyme Activation; D051379:Mice; D008958:Models, Molecular; D010766:Phosphorylation / Q000502:physiology; D011485:Protein Binding; D012694:Serine / Q000378:metabolism; D056564:Sirtuin 1; D037761:Sirtuins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013912:Threonine / Q000378:metabolism
chemical_list: D019926:Cyclin B; D016213:Cyclins; D013912:Threonine; D012694:Serine; D016203:CDC2 Protein Kinase; D018844:Cyclin-Dependent Kinases; C470545:Sirt1 protein, mouse; D056564:Sirtuin 1; D037761:Sirtuins