Blockade of MCU-Mediated Ca 2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation

Dhanendra Tomar, Fabián Jaña, Zhiwei Dong, William J. Quinn, Pooja Jadiya, Sarah L. Breves, Cassidy C. Daw, Subramanya Srikantan, Shanmughapriya Santhanam, Neeharika Nemani, Edmund Carvalho, Aparna Tripathi, Alison M. Worth, Xueqian Zhang, Roshanak Razmpour, Ajay Seelam, Stephen Rhode, Anuj V. Mehta, Michael Murray, Daniel SladeServio H. Ramirez, Prashant Mishra, Glenn S. Gerhard, Jeffrey Caplan, Luke Norton, Kumar Sharma, Sudarsan Rajan, Darius Balciunas, Dayanjan S. Wijesinghe, Rexford S. Ahima, Joseph A. Baur, Muniswamy Madesh

Research output: Contribution to journalArticle

Abstract

Mitochondrial Ca 2+ uniporter (MCU)-mediated Ca 2+ uptake promotes the buildup of reducing equivalents that fuel oxidative phosphorylation for cellular metabolism. Although MCU modulates mitochondrial bioenergetics, its function in energy homeostasis in vivo remains elusive. Here we demonstrate that deletion of the Mcu gene in mouse liver (MCU Δhep ) and in Danio rerio by CRISPR/Cas9 inhibits mitochondrial Ca 2+ ( m Ca 2+ ) uptake, delays cytosolic Ca 2+ ( c Ca 2+ ) clearance, reduces oxidative phosphorylation, and leads to increased lipid accumulation. Elevated hepatic lipids in MCU Δhep were a direct result of extramitochondrial Ca 2+ -dependent protein phosphatase-4 (PP4) activity, which dephosphorylates AMPK. Loss of AMPK recapitulates hepatic lipid accumulation without changes in MCU-mediated Ca 2+ uptake. Furthermore, reconstitution of active AMPK, or PP4 knockdown, enhances lipid clearance in MCU Δhep hepatocytes. Conversely, gain-of-function MCU promotes rapid m Ca 2+ uptake, decreases PP4 levels, and reduces hepatic lipid accumulation. Thus, our work uncovers an MCU/PP4/AMPK molecular cascade that links Ca 2+ dynamics to hepatic lipid metabolism. Hepatic mitochondrial Ca 2+ shapes bioenergetics and lipid homeostasis. Tomar et al. demonstrate that MCU-mediated c Ca 2+ buffering serves as a crucial step in controlling hepatic fuel metabolism through an MCU/PP4/AMPK molecular cascade. Identification of these molecular signaling events aids in understanding how perturbation of mitochondrial ion homeostasis may contribute to the etiology of metabolic disorders.

Original languageEnglish (US)
Pages (from-to)3709-3725.e7
JournalCell Reports
Volume26
Issue number13
DOIs
StatePublished - Mar 26 2019

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AMP-Activated Protein Kinases
Lipid Metabolism
Lipids
Liver
Homeostasis
Metabolism
Oxidative Phosphorylation
Energy Metabolism
Clustered Regularly Interspaced Short Palindromic Repeats
Gene Deletion
Zebrafish
protein phosphatase 4
Genes
Hepatocytes
Ions

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Tomar, D., Jaña, F., Dong, Z., Quinn, W. J., Jadiya, P., Breves, S. L., ... Madesh, M. (2019). Blockade of MCU-Mediated Ca 2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation Cell Reports, 26(13), 3709-3725.e7. https://doi.org/10.1016/j.celrep.2019.02.107
Tomar, Dhanendra ; Jaña, Fabián ; Dong, Zhiwei ; Quinn, William J. ; Jadiya, Pooja ; Breves, Sarah L. ; Daw, Cassidy C. ; Srikantan, Subramanya ; Santhanam, Shanmughapriya ; Nemani, Neeharika ; Carvalho, Edmund ; Tripathi, Aparna ; Worth, Alison M. ; Zhang, Xueqian ; Razmpour, Roshanak ; Seelam, Ajay ; Rhode, Stephen ; Mehta, Anuj V. ; Murray, Michael ; Slade, Daniel ; Ramirez, Servio H. ; Mishra, Prashant ; Gerhard, Glenn S. ; Caplan, Jeffrey ; Norton, Luke ; Sharma, Kumar ; Rajan, Sudarsan ; Balciunas, Darius ; Wijesinghe, Dayanjan S. ; Ahima, Rexford S. ; Baur, Joseph A. ; Madesh, Muniswamy. / Blockade of MCU-Mediated Ca 2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation In: Cell Reports. 2019 ; Vol. 26, No. 13. pp. 3709-3725.e7.
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abstract = "Mitochondrial Ca 2+ uniporter (MCU)-mediated Ca 2+ uptake promotes the buildup of reducing equivalents that fuel oxidative phosphorylation for cellular metabolism. Although MCU modulates mitochondrial bioenergetics, its function in energy homeostasis in vivo remains elusive. Here we demonstrate that deletion of the Mcu gene in mouse liver (MCU Δhep ) and in Danio rerio by CRISPR/Cas9 inhibits mitochondrial Ca 2+ ( m Ca 2+ ) uptake, delays cytosolic Ca 2+ ( c Ca 2+ ) clearance, reduces oxidative phosphorylation, and leads to increased lipid accumulation. Elevated hepatic lipids in MCU Δhep were a direct result of extramitochondrial Ca 2+ -dependent protein phosphatase-4 (PP4) activity, which dephosphorylates AMPK. Loss of AMPK recapitulates hepatic lipid accumulation without changes in MCU-mediated Ca 2+ uptake. Furthermore, reconstitution of active AMPK, or PP4 knockdown, enhances lipid clearance in MCU Δhep hepatocytes. Conversely, gain-of-function MCU promotes rapid m Ca 2+ uptake, decreases PP4 levels, and reduces hepatic lipid accumulation. Thus, our work uncovers an MCU/PP4/AMPK molecular cascade that links Ca 2+ dynamics to hepatic lipid metabolism. Hepatic mitochondrial Ca 2+ shapes bioenergetics and lipid homeostasis. Tomar et al. demonstrate that MCU-mediated c Ca 2+ buffering serves as a crucial step in controlling hepatic fuel metabolism through an MCU/PP4/AMPK molecular cascade. Identification of these molecular signaling events aids in understanding how perturbation of mitochondrial ion homeostasis may contribute to the etiology of metabolic disorders.",
author = "Dhanendra Tomar and Fabi{\'a}n Ja{\~n}a and Zhiwei Dong and Quinn, {William J.} and Pooja Jadiya and Breves, {Sarah L.} and Daw, {Cassidy C.} and Subramanya Srikantan and Shanmughapriya Santhanam and Neeharika Nemani and Edmund Carvalho and Aparna Tripathi and Worth, {Alison M.} and Xueqian Zhang and Roshanak Razmpour and Ajay Seelam and Stephen Rhode and Mehta, {Anuj V.} and Michael Murray and Daniel Slade and Ramirez, {Servio H.} and Prashant Mishra and Gerhard, {Glenn S.} and Jeffrey Caplan and Luke Norton and Kumar Sharma and Sudarsan Rajan and Darius Balciunas and Wijesinghe, {Dayanjan S.} and Ahima, {Rexford S.} and Baur, {Joseph A.} and Muniswamy Madesh",
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Tomar, D, Jaña, F, Dong, Z, Quinn, WJ, Jadiya, P, Breves, SL, Daw, CC, Srikantan, S, Santhanam, S, Nemani, N, Carvalho, E, Tripathi, A, Worth, AM, Zhang, X, Razmpour, R, Seelam, A, Rhode, S, Mehta, AV, Murray, M, Slade, D, Ramirez, SH, Mishra, P, Gerhard, GS, Caplan, J, Norton, L, Sharma, K, Rajan, S, Balciunas, D, Wijesinghe, DS, Ahima, RS, Baur, JA & Madesh, M 2019, ' Blockade of MCU-Mediated Ca 2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation ', Cell Reports, vol. 26, no. 13, pp. 3709-3725.e7. https://doi.org/10.1016/j.celrep.2019.02.107

Blockade of MCU-Mediated Ca 2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation . / Tomar, Dhanendra; Jaña, Fabián; Dong, Zhiwei; Quinn, William J.; Jadiya, Pooja; Breves, Sarah L.; Daw, Cassidy C.; Srikantan, Subramanya; Santhanam, Shanmughapriya; Nemani, Neeharika; Carvalho, Edmund; Tripathi, Aparna; Worth, Alison M.; Zhang, Xueqian; Razmpour, Roshanak; Seelam, Ajay; Rhode, Stephen; Mehta, Anuj V.; Murray, Michael; Slade, Daniel; Ramirez, Servio H.; Mishra, Prashant; Gerhard, Glenn S.; Caplan, Jeffrey; Norton, Luke; Sharma, Kumar; Rajan, Sudarsan; Balciunas, Darius; Wijesinghe, Dayanjan S.; Ahima, Rexford S.; Baur, Joseph A.; Madesh, Muniswamy.

In: Cell Reports, Vol. 26, No. 13, 26.03.2019, p. 3709-3725.e7.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Blockade of MCU-Mediated Ca 2+ Uptake Perturbs Lipid Metabolism via PP4-Dependent AMPK Dephosphorylation

AU - Tomar, Dhanendra

AU - Jaña, Fabián

AU - Dong, Zhiwei

AU - Quinn, William J.

AU - Jadiya, Pooja

AU - Breves, Sarah L.

AU - Daw, Cassidy C.

AU - Srikantan, Subramanya

AU - Santhanam, Shanmughapriya

AU - Nemani, Neeharika

AU - Carvalho, Edmund

AU - Tripathi, Aparna

AU - Worth, Alison M.

AU - Zhang, Xueqian

AU - Razmpour, Roshanak

AU - Seelam, Ajay

AU - Rhode, Stephen

AU - Mehta, Anuj V.

AU - Murray, Michael

AU - Slade, Daniel

AU - Ramirez, Servio H.

AU - Mishra, Prashant

AU - Gerhard, Glenn S.

AU - Caplan, Jeffrey

AU - Norton, Luke

AU - Sharma, Kumar

AU - Rajan, Sudarsan

AU - Balciunas, Darius

AU - Wijesinghe, Dayanjan S.

AU - Ahima, Rexford S.

AU - Baur, Joseph A.

AU - Madesh, Muniswamy

PY - 2019/3/26

Y1 - 2019/3/26

N2 - Mitochondrial Ca 2+ uniporter (MCU)-mediated Ca 2+ uptake promotes the buildup of reducing equivalents that fuel oxidative phosphorylation for cellular metabolism. Although MCU modulates mitochondrial bioenergetics, its function in energy homeostasis in vivo remains elusive. Here we demonstrate that deletion of the Mcu gene in mouse liver (MCU Δhep ) and in Danio rerio by CRISPR/Cas9 inhibits mitochondrial Ca 2+ ( m Ca 2+ ) uptake, delays cytosolic Ca 2+ ( c Ca 2+ ) clearance, reduces oxidative phosphorylation, and leads to increased lipid accumulation. Elevated hepatic lipids in MCU Δhep were a direct result of extramitochondrial Ca 2+ -dependent protein phosphatase-4 (PP4) activity, which dephosphorylates AMPK. Loss of AMPK recapitulates hepatic lipid accumulation without changes in MCU-mediated Ca 2+ uptake. Furthermore, reconstitution of active AMPK, or PP4 knockdown, enhances lipid clearance in MCU Δhep hepatocytes. Conversely, gain-of-function MCU promotes rapid m Ca 2+ uptake, decreases PP4 levels, and reduces hepatic lipid accumulation. Thus, our work uncovers an MCU/PP4/AMPK molecular cascade that links Ca 2+ dynamics to hepatic lipid metabolism. Hepatic mitochondrial Ca 2+ shapes bioenergetics and lipid homeostasis. Tomar et al. demonstrate that MCU-mediated c Ca 2+ buffering serves as a crucial step in controlling hepatic fuel metabolism through an MCU/PP4/AMPK molecular cascade. Identification of these molecular signaling events aids in understanding how perturbation of mitochondrial ion homeostasis may contribute to the etiology of metabolic disorders.

AB - Mitochondrial Ca 2+ uniporter (MCU)-mediated Ca 2+ uptake promotes the buildup of reducing equivalents that fuel oxidative phosphorylation for cellular metabolism. Although MCU modulates mitochondrial bioenergetics, its function in energy homeostasis in vivo remains elusive. Here we demonstrate that deletion of the Mcu gene in mouse liver (MCU Δhep ) and in Danio rerio by CRISPR/Cas9 inhibits mitochondrial Ca 2+ ( m Ca 2+ ) uptake, delays cytosolic Ca 2+ ( c Ca 2+ ) clearance, reduces oxidative phosphorylation, and leads to increased lipid accumulation. Elevated hepatic lipids in MCU Δhep were a direct result of extramitochondrial Ca 2+ -dependent protein phosphatase-4 (PP4) activity, which dephosphorylates AMPK. Loss of AMPK recapitulates hepatic lipid accumulation without changes in MCU-mediated Ca 2+ uptake. Furthermore, reconstitution of active AMPK, or PP4 knockdown, enhances lipid clearance in MCU Δhep hepatocytes. Conversely, gain-of-function MCU promotes rapid m Ca 2+ uptake, decreases PP4 levels, and reduces hepatic lipid accumulation. Thus, our work uncovers an MCU/PP4/AMPK molecular cascade that links Ca 2+ dynamics to hepatic lipid metabolism. Hepatic mitochondrial Ca 2+ shapes bioenergetics and lipid homeostasis. Tomar et al. demonstrate that MCU-mediated c Ca 2+ buffering serves as a crucial step in controlling hepatic fuel metabolism through an MCU/PP4/AMPK molecular cascade. Identification of these molecular signaling events aids in understanding how perturbation of mitochondrial ion homeostasis may contribute to the etiology of metabolic disorders.

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