Proyectos del Plan Nacional I+D+I con referencias PB94-0129, PB97-1132, BHA 2002-00138, HUM 2006-06250/HISTProyectos de la CAM con referencias 06/0020/1997, 06/0094/1998, 06/0090/2000, 06/0043/2001Programa Consolider-Ingenio 2010 con sigla CSD2007-00058NoMuseo Arqueológico Nacional (Madrid)AbengibrePlato argénteo nº 38.215 . Revers

B\uf6ttcher, M

Buttgereit, A

Cerqua, C

Friedrich, O

Harbauer, Ab

Hashemolhosseini, S.

Heuss, D

Huraskin, D

Kaiser, T

Kravic, B

Litchfield, D

Marber, M

Martin, Ed

Meisinger, C

Mougiakakos, D

Neuhuber, W

Poveda-Huertes, D

Rabalski, Aj

Romanello, V

Rudolf, R

Salviati, L

Sandri, M

Simeone, L

Straubinger, M

V\uf6gtle, Fn

In yeast, Tom22, the central component of the TOMM (translocase of outer mitochondrial membrane) receptor complex, is responsible for the recognition and translocation of synthesized mitochondrial precursor proteins, and its protein kinase CK2-dependent phosphorylation is mandatory for TOMM complex biogenesis and proper mitochondrial protein import. In mammals, the biological function of protein kinase CSNK2/CK2 remains vastly elusive and it is unknown whether CSNK2-dependent phosphorylation of TOMM protein subunits has a similar role as that in yeast. To address this issue, we used a skeletal muscle-specific Csnk2b/Ck2-conditional knockout (cKO) mouse model. Phenotypically, these skeletal muscle Csnk2b cKO mice showed reduced muscle strength and abnormal metabolic activity of mainly oxidative muscle fibers, which point towards mitochondrial dysfunction. Enzymatically, active muscle lysates from skeletal muscle Csnk2b cKO mice phosphorylate murine TOMM22, the mammalian ortholog of yeast Tom22, to a lower extent than lysates prepared from controls. Mechanistically, CSNK2-mediated phosphorylation of TOMM22 changes its binding affinity for mitochondrial precursor proteins. However, in contrast to yeast, mitochondrial protein import seems not to be affected in vitro using mitochondria isolated from muscles of skeletal muscle Csnk2b cKO mice. PINK1, a mitochondrial health sensor that undergoes constitutive import under physiological conditions, accumulates within skeletal muscle Csnk2b cKO fibers and labels abnormal mitochondria for removal by mitophagy as demonstrated by the appearance of mitochondria-containing autophagosomes through electron microscopy. Mitophagy can be normalized by either introduction of a phosphomimetic TOMM22 mutant in cultured myotubes, or by in vivo electroporation of phosphomimetic Tomm22 into muscles of mice. Importantly, transfection of the phosphomimetic Tomm22 mutant in muscle cells with ablated Csnk2b restored their oxygen consumption rate comparable to wild-type levels. In sum, our data show that mammalian CSNK2-dependent phosphorylation of TOMM22 is a critical switch for mitophagy and reveal CSNK2-dependent physiological implications on metabolism, muscle integrity and behavior

Kravic, B.

Harbauer, A.

Romanello, V.

Simeone, L.

Vogtle, F.

Kaiser, T.

Straubinger, M.

Huraskin, D.

Bottcher, M.

Cerqua, C.

Martin, E.

Poveda-Huertes, D.

Buttgereit, A.

Rabalski, A.

Heuss, D.

Rudolf, R.

Friedrich, O.

Litchfield, D.

Marber, M.

Salviati, L.

Mougiakakos, D.

Neuhuber, W.

Sandri, M.

Meisinger, C.

MPG.PuRe

In mammalian skeletal muscle, phosphorylation of TOMM22 by protein kinase CSNK2/CK2 controls mitophagy

In yeast, Tom22 the central component of the TOMM (translocase of outer mitochondrial membrane) receptor complex is responsible for the recognition and translocation of synthesized mitochondrial precursor proteins, and its protein kinase CK2-dependent phosphorylation is mandatory for TOMM complex biogenesis and proper mitochondrial protein import. In mammals, the biological function of protein kinase CSNK2/CK2 remains vastly elusive and it is unknown whether CSNK2-dependent phosphorylation of TOMM protein subunits has a similar role as that in yeast. To address this issue, we used a skeletal muscle-specific Csnk2b/Ck2β-conditional knockout (cKO) mouse model. Phenotypically, these skeletal muscle Csnk2b cKO mice showed reduced muscle strength and abnormal metabolic activity of mainly oxidative muscle fibers, which point towards mitochondrial dysfunction. Enzymatically, active muscle lysates from skeletal muscle Csnk2b cKO mice phosphorylate murine TOMM22, the mammalian ortholog of yeast Tom22, to a lower extent than lysates prepared from controls. Mechanistically, CSNK2-mediated phosphorylation of TOMM22 changes its binding affinity for mitochondrial precursor proteins. However, in contrast to yeast, mitochondrial protein import seems not to be affected in vitro using mitochondria isolated from muscles of skeletal muscle Csnk2b cKO mice. PINK1, a mitochondrial health sensor that undergoes constitutive import under physiological conditions, accumulates within skeletal muscle Csnk2b cKO fibers and labels abnormal mitochondria for removal by mitophagy as demonstrated by the appearance of mitochondria-containing autophagosomes through electron microscopy. Mitophagy can be normalized by either introduction of a phosphomimetic TOMM22 mutant in cultured myotubes, or by in vivo electroporation of phosphomimetic Tomm22 into muscles of mice. Importantly, transfection of the phosphomimetic Tomm22 mutant in muscle cells with ablated Csnk2b restored their oxygen consumption rate comparable to wild-type levels. In sum, our data show that mammalian CSNK2-dependent phosphorylation of TOMM22 is a critical switch for mitophagy and reveal CSNK2-dependent physiological implications on metabolism, muscle integrity and behavior

Kravic B

Harbauer AB

Romanello V

Simeone L

Vögtle FN

Kaiser T

Straubinger M

Huraskin D

Böttcher M

Cerqua C

Martin ED

Poveda-Huertes D

Buttgereit A

Rabalski AJ

Heuss D

Rudolf R

Friedrich O

Litchfield D

Marber M

Salviati L

Mougiakakos D

Neuhuber W

Sandri M

Meisinger C

Hashemolhosseini S.

Archivio istituzionale della ricerca - Università di Padova

In mammalian skeletal muscle, phosphorylation of TOMM22 by protein kinase CSNK2/CK2 controls mitophagy.

© 2018 Taylor and Francis Group, LLC. In yeast, Tom22, the central component of the TOMM (translocase of outer mitochondrial membrane) receptor complex, is responsible for the recognition and translocation of synthesized mitochondrial precursor proteins, and its protein kinase CK2-dependent phosphorylation is mandatory for TOMM complex biogenesis and proper mitochondrial protein import. In mammals, the biological function of protein kinase CSNK2/CK2 remains vastly elusive and it is unknown whether CSNK2-dependent phosphorylation of TOMM protein subunits has a similar role as that in yeast. To address this issue, we used a skeletal muscle-specific Csnk2b/Ck2β-conditional knockout (cKO) mouse model. Phenotypically, these skeletal muscle Csnk2b cKO mice showed reduced muscle strength and abnormal metabolic activity of mainly oxidative muscle fibers, which point towards mitochondrial dysfunction. Enzymatically, active muscle lysates from skeletal muscle Csnk2b cKO mice phosphorylate murine TOMM22, the mammalian ortholog of yeast Tom22, to a lower extent than lysates prepared from controls. Mechanistically, CSNK2-mediated phosphorylation of TOMM22 changes its binding affinity for mitochondrial precursor proteins. However, in contrast to yeast, mitochondrial protein import seems not to be affected in vitro using mitochondria isolated from muscles of skeletal muscle Csnk2b cKO mice. PINK1, a mitochondrial health sensor that undergoes constitutive import under physiological conditions, accumulates within skeletal muscle Csnk2b cKO fibers and labels abnormal mitochondria for removal by mitophagy as demonstrated by the appearance of mitochondria-containing autophagosomes through electron microscopy. Mitophagy can be normalized by either introduction of a phosphomimetic TOMM22 mutant in cultured myotubes, or by in vivo electroporation of phosphomimetic Tomm22 into muscles of mice. Importantly, transfection of the phosphomimetic Tomm22 mutant in muscle cells with ablated Csnk2b restored their oxygen consumption rate comparable to wild-type levels. In sum, our data show that mammalian CSNK2-dependent phosphorylation of TOMM22 is a critical switch for mitophagy and reveal CSNK2-dependent physiological implications on metabolism, muscle integrity and behavior

Kravic, Bojana

Harbauer, Angelika B.

Romanello, Vanina

Simeone, Luca

Vögtle, F. Nora

Kaiser, Tobias

Straubinger, Marion

Huraskin, Danyil

Böttcher, Martin

Cerqua, Cristina

Martin, Eva Denise

Poveda-Huertes, Daniel

Buttgereit, Andreas

Rabalski, Adam J.

Heuss, Dieter

Rudolf, Rüdiger

Friedrich, Oliver

Litchfield, David

Marber, Michael

Salviati, Leonardo

Mougiakakos, Dimitrios

Neuhuber, Winfried

Sandri, Marco

Meisinger, Chris

Hashemolhosseini, Said

Scholarship@Western

English

Bojana Kravic

Angelika B. Harbauer

Vanina Romanello

Luca Simeone

F.-Nora Vögtle

Tobias Kaiser

Marion Straubinger

Danyil Huraskin

Martin Böttcher

Cristina Cerqua

Eva Denise Martin

Daniel Poveda-Huertes

Andreas Buttgereit

Adam J. Rabalski

Dieter Heuss

Rüdiger Rudolf

Oliver Friedrich

David Litchfield

Michael Marber

Leonardo Salviati

Dimitrios Mougiakakos

Winfried Neuhuber

Marco Sandri

Chris Meisinger

Said Hashemolhosseini

Crossref

Autophagy

In yeast, Tom22, the central component of the TOMM (translocase of outer mitochondrial membrane) receptor complex, is responsible for the recognition and translocation of synthesized mitochondrial precursor proteins, and its protein kinase CK2-dependent phosphorylation is mandatory for TOMM complex biogenesis and proper mitochondrial protein import. In mammals, the biological function of protein kinase CSNK2/CK2 remains vastly elusive and it is unknown whether CSNK2-dependent phosphorylation of TOMM protein subunits has a similar role as that in yeast. To address this issue, we used a skeletal muscle-specific Csnk2b/Ck2β-conditional knockout (cKO) mouse model. Phenotypically, these skeletal muscle Csnk2b cKO mice showed reduced muscle strength and abnormal metabolic activity of mainly oxidative muscle fibers, which point towards mitochondrial dysfunction. Enzymatically, active muscle lysates from skeletal muscle Csnk2b cKO mice phosphorylate murine TOMM22, the mammalian ortholog of yeast Tom22, to a lower extent than lysates prepared from controls. Mechanistically, CSNK2-mediated phosphorylation of TOMM22 changes its binding affinity for mitochondrial precursor proteins. However, in contrast to yeast, mitochondrial protein import seems not to be affected in vitro using mitochondria isolated from muscles of skeletal muscle Csnk2b cKO mice. PINK1, a mitochondrial health sensor that undergoes constitutive import under physiological conditions, accumulates within skeletal muscle Csnk2b cKO fibers and labels abnormal mitochondria for removal by mitophagy as demonstrated by the appearance of mitochondria-containing autophagosomes through electron microscopy. Mitophagy can be normalized by either introduction of a phosphomimetic TOMM22 mutant in cultured myotubes, or by in vivo electroporation of phosphomimetic Tomm22 into muscles of mice. Importantly, transfection of the phosphomimetic Tomm22 mutant in muscle cells with ablated Csnk2b restored their oxygen consumption rate comparable to wild-type levels. In sum, our data show that mammalian CSNK2-dependent phosphorylation of TOMM22 is a critical switch for mitophagy and reveal CSNK2-dependent physiological implications on metabolism, muscle integrity and behavior.</p

Bojana Kravic (4609720)

Angelika B. Harbauer (4609696)

Vanina Romanello (4609702)

Luca Simeone (2091757)

F.-Nora Vögtle (3572954)

Tobias Kaiser (4609717)

Marion Straubinger (4609699)

Danyil Huraskin (4609732)

Martin Böttcher (4609738)

Cristina Cerqua (4609711)

Eva Denise Martin (4609708)

Daniel Poveda-Huertes (4609723)

Andreas Buttgereit (4609726)

Adam J. Rabalski (4609705)

Dieter Heuss (4609714)

Rüdiger Rudolf (151449)

Oliver Friedrich (48807)

David Litchfield (4609729)

Michael Marber (4609693)

Leonardo Salviati (362927)

Dimitrios Mougiakakos (149682)

Winfried Neuhuber (219250)

Marco Sandri (84476)

Chris Meisinger (655609)

Said Hashemolhosseini (4609735)

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Proyecto AU

Digital.CSIC