Proteomic investigations of lysine acetylation identify diverse substrates of mitochondrial deacetylase sirt3
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Proteomic investigations of lysine acetylation identify diverse substrates of mitochondrial deacetylase sirt3. / Sol, E-ri Maria; Wagner, Sebastian A; Weinert, Brian T; Kumar, Amit; Kim, Hyun-Seok; Deng, Chu-Xia; Choudhary, Chuna Ram.
In: PloS one, Vol. 7, No. 12, 07.12.2012, p. e50545.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Proteomic investigations of lysine acetylation identify diverse substrates of mitochondrial deacetylase sirt3
AU - Sol, E-ri Maria
AU - Wagner, Sebastian A
AU - Weinert, Brian T
AU - Kumar, Amit
AU - Kim, Hyun-Seok
AU - Deng, Chu-Xia
AU - Choudhary, Chuna Ram
PY - 2012/12/7
Y1 - 2012/12/7
N2 - Lysine acetylation is a posttranslational modification that is dynamically regulated by the activity of acetyltransferases and deacetylases. The human and mouse genomes encode 18 different lysine deacetylases (KDACs) which are key regulators of many cellular processes. Identifying substrates of KDACs and pinpointing the regulated acetylation sites on target proteins may provide important information about the molecular basis of their functions. Here we apply quantitative proteomics to identify endogenous substrates of the mitochondrial deacetylase Sirtuin 3 (Sirt3) by comparing site-specific acetylation in wild-type murine embryonic fibroblasts to Sirt3 knockout cells. We confirm Sirt3-regulated acetylation of several mitochondrial proteins in human cells by comparing acetylation in U2OS cells overexpressing Sirt3 to U2OS cells in which Sirt3 expression was reduced by shRNA. Our data demonstrate that ablation of Sirt3 significantly increases acetylation at dozens of sites on mitochondrial proteins. Substrates of Sirt3 are implicated in various metabolic pathways, including fatty acid metabolism and the tricarboxylic acid cycle. These results imply broader regulatory roles of Sirt3 in the mitochondria by modulating acetylation on diverse substrates. The experimental strategy described here is generic and can be applied to identify endogenous substrates of other lysine deacetylases.
AB - Lysine acetylation is a posttranslational modification that is dynamically regulated by the activity of acetyltransferases and deacetylases. The human and mouse genomes encode 18 different lysine deacetylases (KDACs) which are key regulators of many cellular processes. Identifying substrates of KDACs and pinpointing the regulated acetylation sites on target proteins may provide important information about the molecular basis of their functions. Here we apply quantitative proteomics to identify endogenous substrates of the mitochondrial deacetylase Sirtuin 3 (Sirt3) by comparing site-specific acetylation in wild-type murine embryonic fibroblasts to Sirt3 knockout cells. We confirm Sirt3-regulated acetylation of several mitochondrial proteins in human cells by comparing acetylation in U2OS cells overexpressing Sirt3 to U2OS cells in which Sirt3 expression was reduced by shRNA. Our data demonstrate that ablation of Sirt3 significantly increases acetylation at dozens of sites on mitochondrial proteins. Substrates of Sirt3 are implicated in various metabolic pathways, including fatty acid metabolism and the tricarboxylic acid cycle. These results imply broader regulatory roles of Sirt3 in the mitochondria by modulating acetylation on diverse substrates. The experimental strategy described here is generic and can be applied to identify endogenous substrates of other lysine deacetylases.
KW - Acetylation
KW - Animals
KW - Cell Line
KW - Fibroblasts
KW - Humans
KW - Lysine
KW - Mice
KW - Mice, Knockout
KW - Mitochondria
KW - Proteomics
KW - Sirtuin 3
U2 - 10.1371/journal.pone.0050545
DO - 10.1371/journal.pone.0050545
M3 - Journal article
C2 - 23236377
VL - 7
SP - e50545
JO - PLoS ONE
JF - PLoS ONE
SN - 1932-6203
IS - 12
ER -
ID: 47459836