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PMID: 12186850 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The human silent information regulator (Sir)2 homologue hSIRT3 is a mitochondrial nicotinamide adenine dinucleotide-dependent deacetylase.

The Journal of cell biology ·Vol. 158 ·No. 4 ·2002-08-19 ·Pages 647-57

Schwer B, North BJ, Frye RA, Ott M, Verdin E

Abstract

The yeast silent information regulator (Sir)2 protein links cellular metabolism and transcriptional silencing through its nicotinamide adenine dinucleotide (NAD)-dependent histone deacetylase activity. We report that mitochondria from mammalian cells contain intrinsic NAD-dependent deacetylase activity. This activity is inhibited by the NAD hydrolysis product nicotinamide, but not by trichostatin A, consistent with a class III deacetylase. We identify this deacetylase as the nuclear-encoded human Sir2 homologue hSIRT3, and show that hSIRT3 is located within the mitochondrial matrix. Mitochondrial import of hSIRT3 is dependent on an NH2-terminal amphipathic alpha-helix rich in basic residues. hSIRT3 is proteolytically processed in the mitochondrial matrix to a 28-kD product. This processing can be reconstituted in vitro with recombinant mitochondrial matrix processing peptidase (MPP) and is inhibited by mutation of arginines 99 and 100. The unprocessed form of hSIRT3 is enzymatically inactive and becomes fully activated in vitro after cleavage by MPP. These observations demonstrate the existence of a latent class III deacetylase that becomes catalytically activated upon import into the human mitochondria.

MeSH Terms
Biological Transport Cells, Cultured Enzyme Activation HeLa Cells Histone Deacetylases/metabolism Humans Metalloendopeptidases/metabolism Microscopy, Confocal Mitochondria/enzymology Mitochondrial Proteins/metabolism NAD/metabolism Peptide Hydrolases/metabolism Silent Information Regulator Proteins, Saccharomyces cerevisiae Sirtuin 1 Sirtuin 2 Sirtuin 3 Sirtuins Submitochondrial Particles/enzymology Trans-Activators/metabolism
Chemicals
Mitochondrial Proteins Silent Information Regulator Proteins, Saccharomyces cerevisiae Trans-Activators NAD Peptide Hydrolases Metalloendopeptidases mitochondrial processing peptidase SIR2 protein, S cerevisiae SIRT1 protein, human SIRT3 protein, human Sirtuin 1 Sirtuin 2 Sirtuin 3 Sirtuins Histone Deacetylases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Schwer Bjorn
Gladstone Institute of Virology and Immunology, University of California San Francisco, 365 Vermont Street, San Francisco, CA 94103, USA.
North Brian J
Frye Roy A
Ott Melanie
Verdin Eric
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200205057
Published
2002-08-19
Epub
2002-00-19
Pages
647-57
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2174009
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.200205057 PMC Full Text View journal details
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