Phosphorylation regulates SIRT1 function

Tsutomo Sasaki, Bernhard Maier, Katarzyna D. Koclega, Maksymilian Chruszcz, Wendy Gluba, P. Todd Stukenberg, Wladek Minor, Heidi Scrable

Research output: Contribution to journalArticle

179 Citations (Scopus)

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. Methodology/Principal Findings: 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/Significance: 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.

Original languageEnglish
Article numbere4020
JournalPLoS One
Volume3
Issue number12
DOIs
StatePublished - Dec 24 2008

Fingerprint

Phosphorylation
phosphorylation
cell cycle
Cell Cycle
NAD
Cells
Mass Spectrometry
mice
phosphotransferases (kinases)
Proteins
Phosphotransferases
mass spectrometry
Mass spectrometry
Pharmacology
cells
Sumoylation
Cyclin B
Obese Mice
proteins
dephosphorylation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Sasaki, T., Maier, B., Koclega, K. D., Chruszcz, M., Gluba, W., Stukenberg, P. T., ... Scrable, H. (2008). Phosphorylation regulates SIRT1 function. PLoS One, 3(12), [e4020]. https://doi.org/10.1371/journal.pone.0004020

Phosphorylation regulates SIRT1 function. / Sasaki, Tsutomo; Maier, Bernhard; Koclega, Katarzyna D.; Chruszcz, Maksymilian; Gluba, Wendy; Stukenberg, P. Todd; Minor, Wladek; Scrable, Heidi.

In: PLoS One, Vol. 3, No. 12, e4020, 24.12.2008.

Research output: Contribution to journalArticle

Sasaki, T, Maier, B, Koclega, KD, Chruszcz, M, Gluba, W, Stukenberg, PT, Minor, W & Scrable, H 2008, 'Phosphorylation regulates SIRT1 function', PLoS One, vol. 3, no. 12, e4020. https://doi.org/10.1371/journal.pone.0004020
Sasaki T, Maier B, Koclega KD, Chruszcz M, Gluba W, Stukenberg PT et al. Phosphorylation regulates SIRT1 function. PLoS One. 2008 Dec 24;3(12). e4020. https://doi.org/10.1371/journal.pone.0004020
Sasaki, Tsutomo ; Maier, Bernhard ; Koclega, Katarzyna D. ; Chruszcz, Maksymilian ; Gluba, Wendy ; Stukenberg, P. Todd ; Minor, Wladek ; Scrable, Heidi. / Phosphorylation regulates SIRT1 function. In: PLoS One. 2008 ; Vol. 3, No. 12.
@article{e3cb661c4c514faca0d4d57c508ca9b2,
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. Methodology/Principal Findings: 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/Significance: 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.",
author = "Tsutomo Sasaki and Bernhard Maier and Koclega, {Katarzyna D.} and Maksymilian Chruszcz and Wendy Gluba and Stukenberg, {P. Todd} and Wladek Minor and Heidi Scrable",
year = "2008",
month = "12",
day = "24",
doi = "10.1371/journal.pone.0004020",
language = "English",
volume = "3",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "12",

}

TY - JOUR

T1 - Phosphorylation regulates SIRT1 function

AU - Sasaki, Tsutomo

AU - Maier, Bernhard

AU - Koclega, Katarzyna D.

AU - Chruszcz, Maksymilian

AU - Gluba, Wendy

AU - Stukenberg, P. Todd

AU - Minor, Wladek

AU - Scrable, Heidi

PY - 2008/12/24

Y1 - 2008/12/24

N2 - 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. Methodology/Principal Findings: 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/Significance: 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.

AB - 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. Methodology/Principal Findings: 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/Significance: 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.

UR - http://www.scopus.com/inward/record.url?scp=58149202185&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=58149202185&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0004020

DO - 10.1371/journal.pone.0004020

M3 - Article

VL - 3

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 12

M1 - e4020

ER -