A large number of mitochondrial proteins are subject to reversible lysine acetylation, suggesting that this modification plays a significant role in mitochondrial biology. Three distinct NAD+dependent protein deacetylases, called SIRT3, 4 and 5, have been identified in mitochondria. The PIs of this proposal have extensively collaborated to characterize the phenotype of knockout mice lacking SIRT3. These mice show marked mitochondrial protein acetylation. SIRT3 expression increases during fasting and activates (3-oxidation of fatty acid in the liver. SIRTS directly deacetylates two key mitochondrial enzymes in the fatty acid oxidation pathway and increases their enzymatic activities. Metabolomic analysis of mice lacking SIRTS shows significantly greater levels of acylcarnitines during fasting than wild-type mice. SIRTS-/- mice also show reduced ATP levels and intolerance to cold exposure upon fasting consistent with their fatty acid oxidation defect. When SIRTS-/- mice are placed on a high fat diet, they show accelerated development of a syndrome that closely mimics the human metabolic syndrome: obesity, type II diabetes, lipid abnormalities, steatohepatltis and hepatocellular carcinoma. These findings identify acetylation as a novel regulatory mechanism for mitochondrial metabolism. This proposal will use knockout and transgenic mice for SIRTS, 4 and 5 to further explore the mitochondrial acetylproteome, i.e. the proteins that are reversibly deacetylated by SIRTS, 4 and 5. The metabolome of mice lacking SIRTS, 4 and 5 will be explored in a variety of organs to identify metabolic pathways that are regulated by individual mitochondrial sirtuins. We will further explore the molecular mechanism(s) responsible for the development of type II diabetes in mice lacking SIRTS and its link to disordered lipid metabolism. Finally, we will study the regulation of expression of mitochondrial sirtuins under different physiological conditions and screen a small molecule library for drugs that enhance SIRTS expression. The effect of these drugs on the manifestations ofthe metabolic syndrome will be examined in mice. We anticipate that these experiments will increase our understanding of the role of reversible mitochondrial protein acetylation and its enzymes in metabolic regulation and in type II diabetes.

Public Health Relevance

Mitochondria are key players in the pathogenesis of several metabolic disorders. The recent identification of reversible lysine acetylation on a large number of mitochondrial proteins and the identification of three mitochondrial sirtuin (SIRTS, 4 and 5) suggest that reversible mitochondrial acetylation plays a role in the pathogenesis of metabolic disorders. Understanding the role of SIRT 3, 4, 5 in the pathogenesis of the metabolic syndrome and other metabolic disorders could yield novel therapeutic opportunities.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Resource-Related Research Projects (R24)
Project #
5R24DK085610-02
Application #
8074362
Study Section
Special Emphasis Panel (ZDK1-GRB-W (O7))
Program Officer
Sechi, Salvatore
Project Start
2010-06-01
Project End
2012-06-30
Budget Start
2011-06-01
Budget End
2012-06-30
Support Year
2
Fiscal Year
2011
Total Cost
$1,655,718
Indirect Cost
Name
J. David Gladstone Institutes
Department
Type
DUNS #
099992430
City
San Francisco
State
CA
Country
United States
Zip Code
94158
Carrico, Chris; Meyer, Jesse G; He, Wenjuan et al. (2018) The Mitochondrial Acylome Emerges: Proteomics, Regulation by Sirtuins, and Metabolic and Disease Implications. Cell Metab 27:497-512
Peterson, Brett S; Campbell, Jonathan E; Ilkayeva, Olga et al. (2018) Remodeling of the Acetylproteome by SIRT3 Manipulation Fails to Affect Insulin Secretion or ? Cell Metabolism in the Absence of Overnutrition. Cell Rep 24:209-223.e6
Wei, Lei; Meyer, Jesse G; Schilling, Birgit (2018) Quantification of Site-specific Protein Lysine Acetylation and Succinylation Stoichiometry Using Data-independent Acquisition Mass Spectrometry. J Vis Exp :
Softic, Samir; Gupta, Manoj K; Wang, Guo-Xiao et al. (2018) Divergent effects of glucose and fructose on hepatic lipogenesis and insulin signaling. J Clin Invest 128:1199
Basisty, Nathan; Meyer, Jesse G; Wei, Lei et al. (2018) Simultaneous Quantification of the Acetylome and Succinylome by 'One-Pot' Affinity Enrichment. Proteomics 18:e1800123
Newgard, Christopher B (2017) Metabolomics and Metabolic Diseases: Where Do We Stand? Cell Metab 25:43-56
Newman, John C; Verdin, Eric (2017) ?-Hydroxybutyrate: A Signaling Metabolite. Annu Rev Nutr 37:51-76
Newman, John C; Covarrubias, Anthony J; Zhao, Minghao et al. (2017) Ketogenic Diet Reduces Midlife Mortality and Improves Memory in Aging Mice. Cell Metab 26:547-557.e8
Giles, Daniel A; Moreno-Fernandez, Maria E; Stankiewicz, Traci E et al. (2017) Thermoneutral housing exacerbates nonalcoholic fatty liver disease in mice and allows for sex-independent disease modeling. Nat Med 23:829-838
Meyer, Jesse G; Schilling, Birgit (2017) Clinical applications of quantitative proteomics using targeted and untargeted data-independent acquisition techniques. Expert Rev Proteomics 14:419-429

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