Sirtuins are NAD+-dependent protein deacetylases. They mediate adaptive responses to a variety of stresses, including calorie restriction and metabolic stress. Sirtuin 3 (SIRT3) is localized in the mitochondrial matrix, where it regulates the acetylation levels of metabolic enzymes, including acetyl coenzyme A synthetase 2 (refs 1, 2). Mice lacking both Sirt3 alleles appear phenotypically normal under basal conditions, but show marked hyperacetylation of several mitochondrial proteins3. Here we report that SIRT3 expression is upregulated during fasting in liver and brown adipose tissues. During fasting, livers from mice lacking SIRT3 had higher levels of fatty-acid oxidation intermediate products and triglycerides, associated with decreased levels of fatty-acid oxidation, compared to livers from wild-type mice. Mass spectrometry of mitochondrial proteins shows that long-chain acyl coenzyme A dehydrogenase (LCAD) is hyperacetylated at lysine 42 in the absence of SIRT3. LCAD is deacetylated in wild-type mice under fasted conditions and by SIRT3 in vitro and in vivo; and hyperacetylation of LCAD reduces its enzymatic activity. Mice lacking SIRT3 exhibit hallmarks of fatty-acid oxidation disorders during fasting, including reduced ATP levels and intolerance to cold exposure. These findings identify acetylation as a novel regulatory mechanism for mitochondrial fatty-acid oxidation and demonstrate that SIRT3 modulates mitochondrial intermediary metabolism and fatty-acid use during fasting.
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We thank T. Canavan and J. J. Maher for primary hepatocyte preparation (P30 DK026743); C. Harris, L. Swift and the Mouse Metabolic Phenotyping Center (DK59637) for analysis of plasma and tissue lipid analysis; J. Wong for electron microscopy studies, S. Mihalik and D. Cuebas for synthesis of the 2,6-dimethylheptanoyl-CoA; A.-W. Mohsen for purification of recombinant pig ETF; Y.-C. Si for assistance with adenoviral studies; A. Wilson and J. Carroll for preparation of figures; and G. Howard and S. Ordway for editorial review. This work was supported in part by a Senior Scholarship in Aging from the Elison Medical Foundation to E.V. and by institutional support from the J. David Gladstone Institutes. F.W.A. is an Investigator of the Howard Hughes Medical Institute and recipient of an Ellison Medical Foundation Senior Scholar Award. D.B.L. is supported by a K08 award from the National Institute on Aging and the National Institutes of Health. B.S. is supported by an Ellison Medical Foundation/AFAR Senior Postdoctoral Research Grant. N.B.R. and A.K.S. are supported by National Institutes of Health (NIH) grants PO1 HL068758-06A1, DK019514-29 and R01 DK067509-04.
Author Contributions M.D.H., T.S., E.G., E.J., B.S., C.A.G., C.H., S.B. and A.K.S. performed in vitro, in vivo and biochemical studies; O.R.I., R.D.S. and J.R.B. performed metabolomic studies; D.B.L. and Y.L. performed mass spectrometry studies; M.D.H. and E.V. designed the studies, analysed the data and wrote the manuscript; all other authors reviewed and commented on the manuscript.
The authors declare no competing financial interests.
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Hirschey, M., Shimazu, T., Goetzman, E. et al. SIRT3 regulates mitochondrial fatty-acid oxidation by reversible enzyme deacetylation. Nature 464, 121–125 (2010). https://doi.org/10.1038/nature08778
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