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The regulation of AMPK signaling in a natural state of profound metabolic rate depression

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Abstract

In response to energy stress (and elevated AMP), the AMP-activated protein kinase (AMPK) coordinates the restoration of energy homeostasis. We determined that AMPK is activated in a model system (desert snail Otala lactea) during a physiological state of profound metabolic rate depression (estivation) in the absence of a rise in AMP. Kinetic characterization indicated a strong increase in AMPK activity and phosphorylation in estivation, consistent with an increase in P-Ser428 LKB, an established regulator of AMPK. Accordingly, ~2-fold increases in AMPKα1 protein and activity were observed with LKB1 immunoprecipitates from estivating snails. In vitro studies determined that AMPK in crude extracts was activated in the presence of cGMP and deactivated in conditions that permitted protein phosphatase type-2A (PP2A) activity. Furthermore, AMPKα1 protein and activity increased in PKG immunoprecipitates from estivating tissues, suggesting a novel role for PKG in the regulation of AMPK in vivo. We evaluated several downstream targets of AMPK. Acetyl-CoA carboxylase (ACC) activity was strongly inhibited in estivation, consistent with increased P-Ser79 content, and in vitro stimulation of AMPK negated citrate’s ability to stimulate ACC aggregation. Analysis of other targets revealed a strong decrease in PPARγ-coactivator 1α expression in both tissues, which was related to decreased gluconeogenic protein expression in hepatic tissue, but no changes in mitochondrial biogenesis markers in muscle. We concluded that AMPK activation in O. lactea aids in facilitating the suppression of anabolic pathways, without necessarily activating ATP-generating catabolism.

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Abbreviations

AMPK:

AMP-activated protein kinase

ACC:

Acetyl-CoA carboxylase

PGC1α:

PPARγ-coactivator 1α

PKG:

Protein kinase G

PP2A:

Protein phosphatase type-2A

G6Pase:

Glucose-6-phosphatase

PEPCK:

Phosphoenolpyruvate carboxykinase

NRF1:

Nuclear respiratory factor 1

mtTFA:

Mitochondrial transcription factor A

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Acknowledgments

Thanks are due to J. M. Storey for editorial comment on the manuscript. The research was supported by a discovery grant from the Natural Sciences and Engineering Research Council ( NSERC) of Canada (OPG 6793) to K.B.S., by an NSERC postgraduate scholarship to C.J.R., and by an NSERC USRA scholarship to A.G.G.

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Authors and Affiliations

  1. Department of Molecular Physiology, Vanderbilt University School of Medicine, 710 Robinson Research Building, 2200 Pierce Avenue, Nashville, TN, 37232, USA

    Christopher J. Ramnanan

  2. Institute of Biochemistry and Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada

    David C. McMullen, Amy G. Groom & Kenneth B. Storey

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  1. Christopher J. Ramnanan
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  2. David C. McMullen
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Correspondence to Christopher J. Ramnanan.

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Ramnanan, C.J., McMullen, D.C., Groom, A.G. et al. The regulation of AMPK signaling in a natural state of profound metabolic rate depression. Mol Cell Biochem 335, 91–105 (2010). https://doi.org/10.1007/s11010-009-0246-7

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