Over the last several years, accumulating evidence suggests that reversible acetylation may be a major regulatory mechanism for controlling protein function. Recent proteomic investigations have cataloged the existence of hundreds of acetylated proteins, implicating a potential regulatory role for nearly all facets of cellular biochemistry. Interestingly, metabolic enzymes constitute the largest portion of acetylated proteins. Only in a handful of examples has the functional significance of protein acetylation been revealed. Thus, there is tremendous need to determine the regulatory functions of protein acetylation, both at the level of physiological outcomes and at the level of molecular mechanism. This proposal directly investigates the central hypothesis that reversible acetylation is a major regulatory mechanism for controlling protein function. To accomplish these goals, we will investigate the biochemical and biological functions of sirtuins, NAD+dependent protein deacetylases, which are implicated in genome maintenance, metabolism, cell survival, and lifespan.
The specific aims are focused on demonstrating a direct regulatory role for sirtuins in several major metabolic processes, and elucidating how site-specific acetylation affects enzyme function. Employing mechanistic enzymology, quantitative mass spectrometry, novel high-throughput assays, metabolic pathway analysis, structural biology, enzyme regulation, cell culture models, and mouse genetics, these studies will provide the first comprehensive understanding of the functional significance of reversible protein acetylation. The results have the potential to uncover the prominence of a previously-unknown regulatory mechanism and to transform how we understand metabolic and aged-related diseases.

Public Health Relevance

New evidence suggests that a previously-unknown form of cellular and metabolic regulation exits. This proposal seeks to investigate the functional importance of this regulatory mechanism and the role played by a group of enzymes that are implicated in genome maintenance, metabolism, cell survival, and lifespan. The results have the potential to the transform how we understand metabolic and aged-related diseases, and to generate novel therapeutic opportunities.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM065386-12
Application #
8453444
Study Section
Macromolecular Structure and Function E Study Section (MSFE)
Program Officer
Gerratana, Barbara
Project Start
2003-05-01
Project End
2015-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
12
Fiscal Year
2013
Total Cost
$372,034
Indirect Cost
$122,813
Name
University of Wisconsin Madison
Department
Biochemistry
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
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Hullinger, Rikki; Li, Mi; Wang, Jingxin et al. (2016) Increased expression of AT-1/SLC33A1 causes an autistic-like phenotype in mice by affecting dendritic branching and spine formation. J Exp Med 213:1267-84
Kugel, Sita; Feldman, Jessica L; Klein, Mark A et al. (2015) Identification of and Molecular Basis for SIRT6 Loss-of-Function Point Mutations in Cancer. Cell Rep 13:479-88
Baeza, Josue; Smallegan, Michael J; Denu, John M (2015) Site-specific reactivity of nonenzymatic lysine acetylation. ACS Chem Biol 10:122-8
Feldman, Jessica L; Dittenhafer-Reed, Kristin E; Kudo, Norio et al. (2015) Kinetic and Structural Basis for Acyl-Group Selectivity and NAD(+) Dependence in Sirtuin-Catalyzed Deacylation. Biochemistry 54:3037-50
Dittenhafer-Reed, Kristin E; Richards, Alicia L; Fan, Jing et al. (2015) SIRT3 mediates multi-tissue coupling for metabolic fuel switching. Cell Metab 21:637-46
Sahar, Saurabh; Masubuchi, Satoru; Eckel-Mahan, Kristin et al. (2014) Circadian control of fatty acid elongation by SIRT1 protein-mediated deacetylation of acetyl-coenzyme A synthetase 1. J Biol Chem 289:6091-7
Baeza, Josue; Dowell, James A; Smallegan, Michael J et al. (2014) Stoichiometry of site-specific lysine acetylation in an entire proteome. J Biol Chem 289:21326-38
Prolla, Tomas A; Denu, John M (2014) NAD+ deficiency in age-related mitochondrial dysfunction. Cell Metab 19:178-80

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