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

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

doi:10.1080/15548627.2017.1403716

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

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 SalviatiDimitrios Mougiakakos, Winfried Neuhuber, Marco Sandri, Chris Meisinger, Said Hashemolhosseini^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

46 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	311-335
Number of pages	25
Journal	Autophagy
Volume	14
Issue number	2
DOIs	https://doi.org/10.1080/15548627.2017.1403716
Publication status	Published - 1 Feb 2018

Keywords

CSNK2/CK2
CSNK2B
homeostasis
mitochondria
mitophagy
p62
PINK1
skeletal myopathy
TOMM22

Access to Document

10.1080/15548627.2017.1403716

Cite this

Kravic, B., Harbauer, A. B., Romanello, V., Simeone, L., Vögtle, F. N., Kaiser, T., Straubinger, M., Huraskin, D., Böttcher, M., Cerqua, C., Martin, E. D., Poveda-Huertes, D., Buttgereit, A., Rabalski, A. J., Heuss, D., Rudolf, R., Friedrich, O., Litchfield, D., Marber, M., ... Hashemolhosseini, S. (2018). In mammalian skeletal muscle, phosphorylation of TOMM22 by protein kinase CSNK2/CK2 controls mitophagy. Autophagy, 14(2), 311-335. https://doi.org/10.1080/15548627.2017.1403716

@article{b01bdb1833cb4a62b655159936447151,

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

abstract = "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.",

keywords = "CSNK2/CK2, CSNK2B, homeostasis, mitochondria, mitophagy, p62, PINK1, skeletal myopathy, TOMM22",

author = "Bojana Kravic and Harbauer, {Angelika B.} and Vanina Romanello and Luca Simeone and V{\"o}gtle, {F. Nora} and Tobias Kaiser and Marion Straubinger and Danyil Huraskin and Martin B{\"o}ttcher and Cristina Cerqua and Martin, {Eva Denise} and Daniel Poveda-Huertes and Andreas Buttgereit and Rabalski, {Adam J.} and Dieter Heuss and R{\"u}diger Rudolf and Oliver Friedrich and David Litchfield and Michael Marber and Leonardo Salviati and Dimitrios Mougiakakos and Winfried Neuhuber and Marco Sandri and Chris Meisinger and Said Hashemolhosseini",

year = "2018",

month = feb,

day = "1",

doi = "10.1080/15548627.2017.1403716",

language = "English",

volume = "14",

pages = "311--335",

journal = "Autophagy",

issn = "1554-8627",

publisher = "Landes Bioscience",

number = "2",

}

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 2018, 'In mammalian skeletal muscle, phosphorylation of TOMM22 by protein kinase CSNK2/CK2 controls mitophagy', Autophagy, vol. 14, no. 2, pp. 311-335. https://doi.org/10.1080/15548627.2017.1403716

TY - JOUR

T1 - In mammalian skeletal muscle, phosphorylation of TOMM22 by protein kinase CSNK2/CK2 controls mitophagy

AU - Kravic, Bojana

AU - Harbauer, Angelika B.

AU - Romanello, Vanina

AU - Simeone, Luca

AU - Vögtle, F. Nora

AU - Kaiser, Tobias

AU - Straubinger, Marion

AU - Huraskin, Danyil

AU - Böttcher, Martin

AU - Cerqua, Cristina

AU - Martin, Eva Denise

AU - Poveda-Huertes, Daniel

AU - Buttgereit, Andreas

AU - Rabalski, Adam J.

AU - Heuss, Dieter

AU - Rudolf, Rüdiger

AU - Friedrich, Oliver

AU - Litchfield, David

AU - Marber, Michael

AU - Salviati, Leonardo

AU - Mougiakakos, Dimitrios

AU - Neuhuber, Winfried

AU - Sandri, Marco

AU - Meisinger, Chris

AU - Hashemolhosseini, Said

PY - 2018/2/1

Y1 - 2018/2/1

N2 - 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.

AB - 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.

KW - CSNK2/CK2

KW - CSNK2B

KW - homeostasis

KW - mitochondria

KW - mitophagy

KW - p62

KW - PINK1

KW - skeletal myopathy

KW - TOMM22

UR - http://www.scopus.com/inward/record.url?scp=85041599580&partnerID=8YFLogxK

U2 - 10.1080/15548627.2017.1403716

DO - 10.1080/15548627.2017.1403716

M3 - Article

AN - SCOPUS:85041599580

SN - 1554-8627

VL - 14

SP - 311

EP - 335

JO - Autophagy

JF - Autophagy

IS - 2

ER -

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

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this