PMID:19241369

Citation	Jin, L, Galonek, H, Israelian, K, Choy, W, Morrison, M, Xia, Y, Wang, X, Xu, Y, Yang, Y, Smith, JJ, Hoffmann, E, Carney, DP, Perni, RB, Jirousek, MR, Bemis, JE, Milne, JC, Sinclair, DA and Westphal, CH (2009) Biochemical characterization, localization, and tissue distribution of the longer form of mouse SIRT3. Protein Sci. 18:514-25
Abstract	SIRT3 is a key mitochondrial protein deacetylase proposed to play key roles in regulating mitochondrial metabolism but there has been considerable debate about its actual size, the sequences required for activity, and its subcellular localization. A previously cloned mouse SIRT3 has high sequence similarity with the C-terminus of human SIRT3 but lacks an N-terminal mitochondrial targeting sequence and has no detectable deacetylation activity in vitro. Using 5' rapid amplification of cDNA ends, we cloned the entire sequence of mouse SIRT3, as well as rat and rabbit SIRT3. Importantly, we find that full-length SIRT3 protein localizes exclusively to the mitochondria, in contrast to reports of SIRT3 localization to the nucleus. We demonstrate that SIRT3 has no deacetylation activity in vitro unless the protein is truncated, consistent with human SIRT3. In addition, we determined the inhibition constants and mechanism of action for nicotinamide and a small molecule SIRT3 inhibitor against active mouse SIRT3 and show that the mechanisms are different for the two compounds with respect to peptide substrate and NAD(+). Thus, identification and characterization of the actual SIRT3 sequence should help resolve the debate about the nature of mouse SIRT3 and identify new mechanisms to modulate enzymatic activity.
Links	PubMed PMC2760358 Online version:10.1002/pro.50
Keywords	Amino Acid Sequence; Animals; Base Sequence; Cells, Cultured; Cloning, Molecular; Heterocyclic Compounds with 4 or More Rings/metabolism; Mice; Mitochondria/metabolism; Mitochondrial Proteins/antagonists & inhibitors; Mitochondrial Proteins/chemistry; Mitochondrial Proteins/genetics; Mitochondrial Proteins/metabolism; Molecular Sequence Data; Niacinamide/metabolism; Protein Sorting Signals; Rabbits; Rats; Recombinant Fusion Proteins/antagonists & inhibitors; Recombinant Fusion Proteins/chemistry; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Sequence Alignment; Sirtuin 3; Sirtuins/antagonists & inhibitors; Sirtuins/chemistry; Sirtuins/genetics; Sirtuins/metabolism; Tissue Distribution/genetics
edit table

Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
MOUSE:SIR3	enables	GO:0032041: NAD-dependent histone deacetylase activity (H3-K14 specific)	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
MOUSE:SIR3	acts_upstream_of_or_within	GO:0006476: protein deacetylation	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
MOUSE:SIR3	part_of	GO:0005739: mitochondrion	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete

References

See Help:References for how to manage references in GONUTS.