Journal of Biological Chemistry · 2009 · 165 citations · 48 references
Life SciencesBiochemistryMolecular BiologyEpigeneticsMetabolismProtein TranslationMitochondrial BiochemistryMitochondrial Gene ExpressionMitochondrial MetabolismMitochondrial RibosomesMitochondrial TranslationRibosome BiogenesisHistone DeacetylasesMitochondrial Protein Synthesis InhibitionMitochondrial Protein Synthesis RegulationMitochondrial Ribosomal ProteinsMitochondrial Ribosome FunctionMitochondrial Ribosome RegulationRibosomal Protein SynthesisSirtuin Regulation
A member of the sirtuin family of NAD+-dependent deacetylases, SIRT3, is located in mammalian mitochondria and is important for regulation of mitochondrial metabolism, cell survival, and longevity. In this study, MRPL10 (mitochondrial ribosomal protein L10) was identified as the major acetylated protein in the mitochondrial ribosome. Ribosome-associated SIRT3 was found to be responsible for deacetylation of MRPL10 in an NAD+-dependent manner. We mapped the acetylated Lys residues by tandem mass spectrometry and determined the role of these residues in acetylation of MRPL10 by site-directed mutagenesis. Furthermore, we observed that the increased acetylation of MRPL10 led to an increase in translational activity of mitochondrial ribosomes in Sirt3−/− mice. In a similar manner, ectopic expression and knockdown of SIRT3 in C2C12 cells resulted in the suppression and enhancement of mitochondrial protein synthesis, respectively. Our findings constitute the first evidence for the regulation of mitochondrial protein synthesis by the reversible acetylation of the mitochondrial ribosome and characterize MRPL10 as a novel substrate of the NAD+-dependent deacetylase, SIRT3. A member of the sirtuin family of NAD+-dependent deacetylases, SIRT3, is located in mammalian mitochondria and is important for regulation of mitochondrial metabolism, cell survival, and longevity. In this study, MRPL10 (mitochondrial ribosomal protein L10) was identified as the major acetylated protein in the mitochondrial ribosome. Ribosome-associated SIRT3 was found to be responsible for deacetylation of MRPL10 in an NAD+-dependent manner. We mapped the acetylated Lys residues by tandem mass spectrometry and determined the role of these residues in acetylation of MRPL10 by site-directed mutagenesis. Furthermore, we observed that the increased acetylation of MRPL10 led to an increase in translational activity of mitochondrial ribosomes in Sirt3−/− mice. In a similar manner, ectopic expression and knockdown of SIRT3 in C2C12 cells resulted in the suppression and enhancement of mitochondrial protein synthesis, respectively. Our findings constitute the first evidence for the regulation of mitochondrial protein synthesis by the reversible acetylation of the mitochondrial ribosome and characterize MRPL10 as a novel substrate of the NAD+-dependent deacetylase, SIRT3. IntroductionMitochondria produce over 90% of the energy used by mammalian cells through the process of oxidative phosphorylation. Reversible acetylation regulates many biological processes, including mitochondrial energy metabolism (1.Kim S.C. Sprung R. Chen Y. Xu Y. Ball H. Pei J. Cheng T. Kho Y. Xiao H. Xiao L. Grishin N.V. White M. Yang X.J. Zhao Y. Mol. Cell. 2006; 23: 607-618Abstract Full Text Full Text PDF PubMed Scopus (1203) Google Scholar, 2.Jackson P.J. Harris D.A. Biochem. J. 1986; 235: 577-583Crossref PubMed Scopus (24) Google Scholar, 3.Hallows W.C. Lee S. Denu J.M. Proc. Natl. Acad. Sci. U.S.A. 2006; 103: 10230-10235Crossref PubMed Scopus (639) Google Scholar, 4.Dinardo M.M. Musicco C. Fracasso F. Milella F. Gadaleta M.N. Gadaleta G. Cantatore P. Biochem. Biophys. Res. Commun. 2003; 301: 187-191Crossref PubMed Scopus (51) Google Scholar, 5.Gerhart-Hines Z. Rodgers J.T. Bare O. Lerin C. Kim S.H. Mostoslavsky R. Alt F.W. Wu Z. Puigserver P. EMBO J. 2007; 26: 1913-1923Crossref PubMed Scopus (988) Google Scholar, 6.Schwer B. Bunkenborg J. Verdin R.O. Andersen J.S. Verdin E. Proc. Natl. Acad. Sci. U.S.A. 2006; 103: 10224-10229Crossref PubMed Scopus (558) Google Scholar). Although the enzymes involved in the acetylation of mitochondrial proteins are not known, members of the class III histone deacetylases (sirtuins), SIRT3, SIRT4, and SIRT5, have been found to reside in mitochondria (6.Schwer B. Bunkenborg J. Verdin R.O. Andersen J.S. Verdin E. Proc. Natl. Acad. Sci. U.S.A. 2006; 103: 10224-10229Crossref PubMed Scopus (558) Google Scholar, 7.Michishita E. Park J.Y. Burneskis J.M. Barrett J.C. Horikawa I. Mol. Biol. Cell. 2005; 16: 4623-4635Crossref PubMed Scopus (1047) Google Scholar, 8.Onyango P. Celic I. McCaffery J.M. Boeke J.D. Feinberg A.P. Proc. Natl. Acad. Sci. U.S.A. 2002; 99: 13653-13658Crossref PubMed Scopus (429) Google Scholar). Sirtuins are homologs of the yeast SIR2 (silent mating type information regulation 2) gene and use NAD+ as a cosubstrate (9.Imai S. Armstrong C.M. Kaeberlein M. Guarente L. Nature. 2000; 403: 795-800Crossref PubMed Scopus (2737) Google Scholar, 10.Smith J.S. Brachmann C.B. Celic I. Kenna M.A. Muhammad S. Starai V.J. Avalos J.L. Escalante-Semerena J.C. Grubmeyer C. Wolberger C. Boeke J.D. Proc. Natl. Acad. Sci. U.S.A. 2000; 97: 6658-6663Crossref PubMed Scopus (613) Google Scholar, 11.Landry J. Sutton A. Tafrov S.T. Heller R.C. Stebbins J. Pillus L. Sternglanz R. Proc. Natl. Acad. Sci. U.S.A. 2000; 97: 5807-5811Crossref PubMed Scopus (805) Google Scholar). Both SIRT3 and SIRT4 are required to maintain cell survival after genotoxic stress in a NAD+-dependent manner (12.Rose G. Dato S. Altomare K. Bellizzi D. Garasto S. Greco V. Passarino G. Feraco E. Mari V. Barbi C. BonaFe M. Franceschi C. Tan Q. Boiko S. Yashin A.I. De Benedictis G. Exp. Gerontol. 2003; 38: 1065-1070Crossref PubMed Scopus (240) Google Scholar, 13.Yang H. Yang T. Baur J.A. Perez E. Matsui T. Carmona J.J. Lamming D.W. Souza-Pinto N.C. Bohr V.A. Rosenzweig A. de Cabo R. Sauve A.A. Sinclair D.A. Cell. 2007; 130: 1095-1107Abstract Full Text Full Text PDF PubMed Scopus (759) Google Scholar). Genetic variations in the human Sirt3 gene have also been linked to longevity (12.Rose G. Dato S. Altomare K. Bellizzi D. Garasto S. Greco V. Passarino G. Feraco E. Mari V. Barbi C. BonaFe M. Franceschi C. Tan Q. Boiko S. Yashin A.I. De Benedictis G. Exp. Gerontol. 2003; 38: 1065-1070Crossref PubMed Scopus (240) Google Scholar, 13.Yang H. Yang T. Baur J.A. Perez E. Matsui T. Carmona J.J. Lamming D.W. Souza-Pinto N.C. Bohr V.A. Rosenzweig A. de Cabo R. Sauve A.A. Sinclair D.A. Cell. 2007; 130: 1095-1107Abstract Full Text Full Text PDF PubMed Scopus (759) Google Scholar). We have previously shown that SIRT3 expression in adipose tissue is increased by caloric restriction and cold exposure (1.Kim S.C. Sprung R. Chen Y. Xu Y. Ball H. Pei J. Cheng T. Kho Y. Xiao H. Xiao L. Grishin N.V. White M. Yang X.J. Zhao Y. Mol. Cell. 2006; 23: 607-618Abstract Full Text Full Text PDF PubMed Scopus (1203) Google Scholar, 14.Shi T. Wang F. Stieren E. Tong Q. J. Biol. Chem. 2005; 280: 13560-13567Abstract Full Text Full Text PDF PubMed Scopus (566) Google Scholar). Mitochondrial acetyl-CoA synthetase 2 and glutamate dehydrogenase are the two key metabolic enzymes regulated through deacetylation by SIRT3 (3.Hallows W.C. Lee S. Denu J.M. Proc. Natl. Acad. Sci. U.S.A. 2006; 103: 10230-10235Crossref PubMed Scopus (639) Google Scholar, 6.Schwer B. Bunkenborg J. Verdin R.O. Andersen J.S. Verdin E. Proc. Natl. Acad. Sci. U.S.A. 2006; 103: 10224-10229Crossref PubMed Scopus (558) Google Scholar, 15.Lombard D.B. Alt F.W. Cheng H.L. Bunkenborg J. Streeper R.S. Mostoslavsky R. Kim J. Yancopoulos G. Valenzuela D. Murphy A. Yang Y. Chen Y. Hirschey M.D. Bronson R.T. Haigis M. Guarente L.P. Farese Jr., R.V. Weissman S. Verdin E. Schwer B. Mol. Cell. Biol. 2007; 27: 8807-8814Crossref PubMed Scopus (949) Google Scholar). Thus, SIRT3 and SIRT4 modulate mitochondrial function in response to its [NADH]/[NAD+] ratio by regulating the activity of key metabolic enzymes.In addition to metabolic enzymes, nucleus-encoded subunits of the electron transport chain complexes were found to be acetylated (1.Kim S.C. Sprung R. Chen Y. Xu Y. Ball H. Pei J. Cheng T. Kho Y. Xiao H. Xiao L. Grishin N.V. White M. Yang X.J. Zhao Y. Mol. Cell. 2006; 23: 607-618Abstract Full Text Full Text PDF PubMed Scopus (1203) Google Scholar). In fact, Complex I subunit NDUFA9 is a SIRT3 substrate, and acetylation/deacetylation of Complex I is proposed to regulate and maintain basal ATP levels in mammalian mitochondria (16.Ahn B.H. Kim H.S. Song S. Lee I.H. Liu J. Vassilopoulos A. Deng C.X. Finkel T. Proc. Natl. Acad. Sci. U.S.A. 2008; 105: 14447-14452Crossref PubMed Scopus (971) Google Scholar).Thirteen of the essential protein components of the electron transport chain, as well as ATP synthase, are the products of genes present in mitochondrial DNA. The synthesis of these proteins is carried out by mitochondrial ribosomes within this organelle. We and others have previously identified 77 mammalian mitochondrial ribosomal proteins, of which 29 are in the small subunit and 48 are in the large subunit (17.O'Brien T.W. Fiesler S.E. Denslow N.D. Thiede B. Wittmann-Liebold B. Mougey E.B. Sylvester J.E. Graack H.R. J. Biol. Chem. 1999; 274: 36043-36051Abstract Full Text Full Text PDF PubMed Scopus (30) Google Scholar, 18.Koc E.C. Burkhart W. Blackburn K. Moseley A. Koc H. Spremulli L.L. J. Biol. Chem. 2000; 275: 32585-32591Abstract Full Text Full Text PDF PubMed Scopus (64) Google Scholar, 19.Cavdar Koc E. Burkhart W. Blackburn K. Moseley A. Spremulli L.L. J. Biol. Chem. 2001; 276: 19363-19374Abstract Full Text Full Text PDF PubMed Scopus (185) Google Scholar, 20.Suzuki T. Terasaki M. Takemoto-Hori C. Hanada T. Ueda T. Wada A. Watanabe K. J. Biol. Chem. 2001; 276: 21724-21736Abstract Full Text Full Text PDF PubMed Scopus (101) Google Scholar, 21.Koc E.C. Burkhart W. Blackburn K. Moyer M.B. Schlatzer D.M. Moseley A. Spremulli L.L. J. Biol. Chem. 2001; 276: 43958-43969Abstract Full Text Full Text PDF PubMed Scopus (199) Google Scholar). About half of these proteins have homologs in bacterial ribosomes, whereas the remainders represent new classes of ribosomal proteins. However, we have observed that the functional core of the mitochondrial ribosome, essential for protein synthesis, was conserved in the cryoelectron microscopy reconstruction studies (22.Sharma M.R. Koc E.C. Datta P.P. Booth T.M. Spremulli L.L. Agrawal R.K. Cell. 2003; 115: 97-108Abstract Full Text Full Text PDF PubMed Scopus (283) Google Scholar).Mammalian mitochondrial ribosomal proteins are all nucleus-encoded, and some of them have been mapped to regions associated with disorders of mitochondrial energy metabolism (23.O'Brien T.W. O'Brien B.J. Norman R.A. Gene. 2005; 354: 147-151Crossref PubMed Scopus (67) Google Scholar). Alterations in expression levels and mutations of these ribosomal proteins affect mitochondrial protein synthesis, cell growth, and apoptosis (24.Miller C. Saada A. Shaul N. Shabtai N. Ben-Shalom E. Shaag A. Hershkovitz E. Elpeleg O. Ann. Neurol. 2004; 56: 734-738Crossref PubMed Scopus (183) Google Scholar, 25.Kissil J.L. Cohen O. Raveh T. Kimchi A. EMBO J. 1999; 18: 353-362Crossref PubMed Scopus (96) Google Scholar, 26.Chintharlapalli S.R. Jasti M. Malladi S. Parsa K.V. Ballestero R.P. González-García M. J. Cell Biochem. 2005; 94: 611-626Crossref PubMed Scopus (36) Google Scholar, 27.Yoo Y.A. Kim M.J. Park J.K. Chung Y.M. Lee J.H. Chi S.G. Kim J.S. Yoo Y.D. Mol. Cell Biol. 2005; 25: 6603-6616Crossref PubMed Scopus (80) Google Scholar, 28.Miller J.L. Koc H. Koc E.C. Protein Sci. 2008; 17: 251-260Crossref PubMed Scopus (34) Google Scholar). Some of the ribosomal proteins with bacterial homologs, such as MRPS12, MRPS16, and MRPL12, have been shown to be essential to support protein synthesis in mitochondria (24.Miller C. Saada A. Shaul N. Shabtai N. Ben-Shalom E. Shaag A. Hershkovitz E. Elpeleg O. Ann. Neurol. 2004; 56: 734-738Crossref PubMed Scopus (183) Google Scholar, 29.Wang Z. Cotney J. Shadel G.S. J. Biol. Chem. 2007; 282: 12610-12618Abstract Full Text Full Text PDF PubMed Scopus (77) Google Scholar, 30.Emdadul Haque M. Grasso D. Miller C. Spremulli L.L. Saada A. Mitochondrion. 2008; 8: 254-261Crossref PubMed Scopus (53) Google Scholar, 31.Zanotto E. Lehtonen V. Jacobs H.T. Biochim. Biophys. Acta. 2008; 1783: 2352-2362Crossref PubMed Scopus (11) Google Scholar).In the present study, we demonstrate for the first time the acetylation of a mitochondrial ribosomal protein, MRPL10 (mitochondrial ribosomal protein L10), and its deacetylation by the NAD+-dependent deacetylase SIRT3. Using various biochemical and proteomics techniques, we also show that SIRT3 interacts with the mitochondrial ribosome. We propose that mitochondrial protein synthesis is regulated by reversible acetylation of MRPL10 and that the NAD+-dependent SIRT3 stimulates deacetylation of MRPL10, consequently regulating protein synthesis in mammalian mitochondria. IntroductionMitochondria produce over 90% of the energy used by mammalian cells through the process of oxidative phosphorylation. Reversible acetylation regulates many biological processes, including mitochondrial energy metabolism (1.Kim S.C. Sprung R. Chen Y. Xu Y. Ball H. Pei J. Cheng T. Kho Y. Xiao H. Xiao L. Grishin N.V. White M. Yang X.J. Zhao Y. Mol. Cell. 2006; 23: 607-618Abstract Full Text Full Text PDF PubMed Scopus (1203) Google Scholar, 2.Jackson P.J. Harris D.A. Biochem. J. 1986; 235: 577-583Crossref PubMed Scopus (24) Google Scholar, 3.Hallows W.C. Lee S. Denu J.M. Proc. Natl. Acad. Sci. U.S.A. 2006; 103: 10230-10235Crossref PubMed Scopus (639) Google Scholar, 4.Dinardo M.M. Musicco C. Fracasso F. Milella F. Gadaleta M.N. Gadaleta G. Cantatore P. Biochem. Biophys. Res. Commun. 2003; 301: 187-191Crossref PubMed Scopus (51) Google Scholar, 5.Gerhart-Hines Z. Rodgers J.T. Bare O. Lerin C. Kim S.H. Mostoslavsky R. Alt F.W. Wu Z. Puigserver P. EMBO J. 2007; 26: 1913-1923Crossref PubMed Scopus (988) Google Scholar, 6.Schwer B. Bunkenborg J. Verdin R.O. Andersen J.S. Verdin E. Proc. Natl. Acad. Sci. U.S.A. 2006; 103: 10224-10229Crossref PubMed Scopus (558) Google Scholar). Although the enzymes involved in the acetylation of mitochondrial proteins are not known, members of the class III histone deacetylases (sirtuins), SIRT3, SIRT4, and SIRT5, have been found to reside in mitochondria (6.Schwer B. Bunkenborg J. Verdin R.O. Andersen J.S. Verdin E. Proc. Natl. Acad. Sci. U.S.A. 2006; 103: 10224-10229Crossref PubMed Scopus (558) Google Scholar, 7.Michishita E. Park J.Y. Burneskis J.M. Barrett J.C. Horikawa I. Mol. Biol. Cell. 2005; 16: 4623-4635Crossref PubMed Scopus (1047) Google Scholar, 8.Onyango P. Celic I. McCaffery J.M. Boeke J.D. Feinberg A.P. Proc. Natl. Acad. Sci. U.S.A. 2002; 99: 13653-13658Crossref PubMed Scopus (429) Google Scholar). Sirtuins are homologs of the yeast SIR2 (silent mating type information regulation 2) gene and use NAD+ as a cosubstrate (9.Imai S. Armstrong C.M. Kaeberlein M. Guarente L. Nature. 2000; 403: 795-800Crossref PubMed Scopus (2737) Google Scholar, 10.Smith J.S. Brachmann C.B. Celic I. Kenna M.A. Muhammad S. Starai V.J. Avalos J.L. Escalante-Semerena J.C. Grubmeyer C. Wolberger C. Boeke J.D. Proc. Natl. Acad. Sci. U.S.A. 2000; 97: 6658-6663Crossref PubMed Scopus (613) Google Scholar, 11.Landry J. Sutton A. Tafrov S.T. Heller R.C. Stebbins J. Pillus L. Sternglanz R. Proc. Natl. Acad. Sci. U.S.A. 2000; 97: 5807-5811Crossref PubMed Scopus (805) Google Scholar). Both SIRT3 and SIRT4 are required to maintain cell survival after genotoxic stress in a NAD+-dependent manner (12.Rose G. Dato S. Altomare K. Bellizzi D. Garasto S. Greco V. Passarino G. Feraco E. Mari V. Barbi C. BonaFe M. Franceschi C. Tan Q. Boiko S. Yashin A.I. De Benedictis G. Exp. Gerontol. 2003; 38: 1065-1070Crossref PubMed Scopus (240) Google Scholar, 13.Yang H. Yang T. Baur J.A. Perez E. Matsui T. Carmona J.J. Lamming D.W. Souza-Pinto N.C. Bohr V.A. Rosenzweig A. de Cabo R. Sauve A.A. Sinclair D.A. Cell. 2007; 130: 1095-1107Abstract Full Text Full Text PDF PubMed Scopus (759) Google Scholar). Genetic variations in the human Sirt3 gene have also been linked to longevity (12.Rose G. Dato S. Altomare K. Bellizzi D. Garasto S. Greco V. Passarino G. Feraco E. Mari V. Barbi C. BonaFe M. Franceschi C. Tan Q. Boiko S. Yashin A.I. De Benedictis G. Exp. Gerontol. 2003; 38: 1065-1070Crossref PubMed Scopus (240) Google Scholar, 13.Yang H. Yang T. Baur J.A. Perez E. Matsui T. Carmona J.J. Lamming D.W. Souza-Pinto N.C. Bohr V.A. Rosenzweig A. de Cabo R. Sauve A.A. Sinclair D.A. Cell. 2007; 130: 1095-1107Abstract Full Text Full Text PDF PubMed Scopus (759) Google Scholar). We have previously shown that SIRT3 expression in adipose tissue is increased by caloric restriction and cold exposure (1.Kim S.C. Sprung R. Chen Y. Xu Y. Ball H. Pei J. Cheng T. Kho Y. Xiao H. Xiao L. Grishin N.V. White M. Yang X.J. Zhao Y. Mol. Cell. 2006; 23: 607-618Abstract Full Text Full Text PDF PubMed Scopus (1203) Google Scholar, 14.Shi T. Wang F. Stieren E. Tong Q. J. Biol. Chem. 2005; 280: 13560-13567Abstract Full Text Full Text PDF PubMed Scopus (566) Google Scholar). Mitochondrial acetyl-CoA synthetase 2 and glutamate dehydrogenase are the two key metabolic enzymes regulated through deacetylation by SIRT3 (3.Hallows W.C. Lee S. Denu J.M. Proc. Natl. Acad. Sci. U.S.A. 2006; 103: 10230-10235Crossref PubMed Scopus (639) Google Scholar, 6.Schwer B. Bunkenborg J. Verdin R.O. Andersen J.S. Verdin E. Proc. Natl. Acad. Sci. U.S.A. 2006; 103: 10224-10229Crossref PubMed Scopus (558) Google Scholar, 15.Lombard D.B. Alt F.W. Cheng H.L. Bunkenborg J. Streeper R.S. Mostoslavsky R. Kim J. Yancopoulos G. Valenzuela D. Murphy A. Yang Y. Chen Y. Hirschey M.D. Bronson R.T. Haigis M. Guarente L.P. Farese Jr., R.V. Weissman S. Verdin E. Schwer B. Mol. Cell. Biol. 2007; 27: 8807-8814Crossref PubMed Scopus (949) Google Scholar). Thus, SIRT3 and SIRT4 modulate mitochondrial function in response to its [NADH]/[NAD+] ratio by regulating the activity of key metabolic enzymes.In addition to metabolic enzymes, nucleus-encoded subunits of the electron transport chain complexes were found to be acetylated (1.Kim S.C. Sprung R. Chen Y. Xu Y. Ball H. Pei J. Cheng T. Kho Y. Xiao H. Xiao L. Grishin N.V. White M. Yang X.J. Zhao Y. Mol. Cell. 2006; 23: 607-618Abstract Full Text Full Text PDF PubMed Scopus (1203) Google Scholar). In fact, Complex I subunit NDUFA9 is a SIRT3 substrate, and acetylation/deacetylation of Complex I is proposed to regulate and maintain basal ATP levels in mammalian mitochondria (16.Ahn B.H. Kim H.S. Song S. Lee I.H. Liu J. Vassilopoulos A. Deng C.X. Finkel T. Proc. Natl. Acad. Sci. U.S.A. 2008; 105: 14447-14452Crossref PubMed Scopus (971) Google Scholar).Thirteen of the essential protein components of the electron transport chain, as well as ATP synthase, are the products of genes present in mitochondrial DNA. The synthesis of these proteins is carried out by mitochondrial ribosomes within this organelle. We and others have previously identified 77 mammalian mitochondrial ribosomal proteins, of which 29 are in the small subunit and 48 are in the large subunit (17.O'Brien T.W. Fiesler S.E. Denslow N.D. Thiede B. Wittmann-Liebold B. Mougey E.B. Sylvester J.E. Graack H.R. J. Biol. Chem. 1999; 274: 36043-36051Abstract Full Text Full Text PDF PubMed Scopus (30) Google Scholar, 18.Koc E.C. Burkhart W. Blackburn K. Moseley A. Koc H. Spremulli L.L. J. Biol. Chem. 2000; 275: 32585-32591Abstract Full Text Full Text PDF PubMed Scopus (64) Google Scholar, 19.Cavdar Koc E. Burkhart W. Blackburn K. Moseley A. Spremulli L.L. J. Biol. Chem. 2001; 276: 19363-19374Abstract Full Text Full Text PDF PubMed Scopus (185) Google Scholar, 20.Suzuki T. Terasaki M. Takemoto-Hori C. Hanada T. Ueda T. Wada A. Watanabe K. J. Biol. Chem. 2001; 276: 21724-21736Abstract Full Text Full Text PDF PubMed Scopus (101) Google Scholar, 21.Koc E.C. Burkhart W. Blackburn K. Moyer M.B. Schlatzer D.M. Moseley A. Spremulli L.L. J. Biol. Chem. 2001; 276: 43958-43969Abstract Full Text Full Text PDF PubMed Scopus (199) Google Scholar). About half of these proteins have homologs in bacterial ribosomes, whereas the remainders represent new classes of ribosomal proteins. However, we have observed that the functional core of the mitochondrial ribosome, essential for protein synthesis, was conserved in the cryoelectron microscopy reconstruction studies (22.Sharma M.R. Koc E.C. Datta P.P. Booth T.M. Spremulli L.L. Agrawal R.K. Cell. 2003; 115: 97-108Abstract Full Text Full Text PDF PubMed Scopus (283) Google Scholar).Mammalian mitochondrial ribosomal proteins are all nucleus-encoded, and some of them have been mapped to regions associated with disorders of mitochondrial energy metabolism (23.O'Brien T.W. O'Brien B.J. Norman R.A. Gene. 2005; 354: 147-151Crossref PubMed Scopus (67) Google Scholar). Alterations in expression levels and mutations of these ribosomal proteins affect mitochondrial protein synthesis, cell growth, and apoptosis (24.Miller C. Saada A. Shaul N. Shabtai N. Ben-Shalom E. Shaag A. Hershkovitz E. Elpeleg O. Ann. Neurol. 2004; 56: 734-738Crossref PubMed Scopus (183) Google Scholar, 25.Kissil J.L. Cohen O. Raveh T. Kimchi A. EMBO J. 1999; 18: 353-362Crossref PubMed Scopus (96) Google Scholar, 26.Chintharlapalli S.R. Jasti M. Malladi S. Parsa K.V. Ballestero R.P. González-García M. J. Cell Biochem. 2005; 94: 611-626Crossref PubMed Scopus (36) Google Scholar, 27.Yoo Y.A. Kim M.J. Park J.K. Chung Y.M. Lee J.H. Chi S.G. Kim J.S. Yoo Y.D. Mol. Cell Biol. 2005; 25: 6603-6616Crossref PubMed Scopus (80) Google Scholar, 28.Miller J.L. Koc H. Koc E.C. Protein Sci. 2008; 17: 251-260Crossref PubMed Scopus (34) Google Scholar). Some of the ribosomal proteins with bacterial homologs, such as MRPS12, MRPS16, and MRPL12, have been shown to be essential to support protein synthesis in mitochondria (24.Miller C. Saada A. Shaul N. Shabtai N. Ben-Shalom E. Shaag A. Hershkovitz E. Elpeleg O. Ann. Neurol. 2004; 56: 734-738Crossref PubMed Scopus (183) Google Scholar, 29.Wang Z. Cotney J. Shadel G.S. J. Biol. Chem. 2007; 282: 12610-12618Abstract Full Text Full Text PDF PubMed Scopus (77) Google Scholar, 30.Emdadul Haque M. Grasso D. Miller C. Spremulli L.L. Saada A. Mitochondrion. 2008; 8: 254-261Crossref PubMed Scopus (53) Google Scholar, 31.Zanotto E. Lehtonen V. Jacobs H.T. Biochim. Biophys. Acta. 2008; 1783: 2352-2362Crossref PubMed Scopus (11) Google Scholar).In the present study, we demonstrate for the first time the acetylation of a mitochondrial ribosomal protein, MRPL10 (mitochondrial ribosomal protein L10), and its deacetylation by the NAD+-dependent deacetylase SIRT3. Using various biochemical and proteomics techniques, we also show that SIRT3 interacts with the mitochondrial ribosome. We propose that mitochondrial protein synthesis is regulated by reversible acetylation of MRPL10 and that the NAD+-dependent SIRT3 stimulates deacetylation of MRPL10, consequently regulating protein synthesis in mammalian mitochondria.
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Substrate and Functional Diversity of Lysine Acetylation Revealed by a Proteomics Survey
Sung Chan Kim, Robert W. Sprung, Yue Chen et al. · Molecular Cell · 2006 · 1.5K citations · Full text