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MondoA:MLX complex regulates glucose-dependent gene expression and links to circadian rhythm in liver and brain of the freeze-tolerant wood frog, Rana sylvatica

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Abstract

The wood frog, Rana sylvatica, is one of only a few vertebrate species that display natural freeze tolerance. Frogs survive the freezing of about two-thirds of their body water as extracellular ice over the winter months. Multiple adaptations support freeze tolerance including metabolic rate depression and the production of huge amounts of glucose (often 200 mM or more) as a cryoprotectant that protects cells from freeze damage. To understand how high glucose levels affect gene expression, we studied MondoA, a glucose sensing transcription factor, and its partner MLX (Max-like protein) to assess their ability to modulate the expression of genes involved in glucose metabolism and circadian rhythm. Wood frog liver and brain tissues were analyzed, assessing protein levels, nuclear distribution, and DNA binding activity of MondoA:MLX during freezing (24 h at − 2.5 °C) and subsequent thawing (8 h returned to 5 °C), as compared with 5 °C controls. Downstream targets of MondoA:MLX were also evaluated: TXNIP (thioredoxin interacting protein), ARRDC4 (arrestin domain containing 4), HK-2 (hexokinase-2), PFKFB-3 (6-phosphofructo-2-kinase isozyme 3) and KLF-10 (Kruppel-like factor-10). Both KLF-10 and PFKFB-3 are also involved in circadian dependant regulation which was also explored in the current study via analysis of BMAL-1 (aryl hydrocarbon receptor nuclear translocator-like protein 1) and CLOCK (circadian locomotor output cycles kaput) proteins. Our data establish the MondoA-MLX complex as active under the hyperglycemic conditions in liver to regulate glucose metabolism and may also link to circadian rhythm in liver via KLF-10 and PFKFB-3 but not in brain.

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Acknowledgements

Research was supported by a Discovery Grant to KBS from the Natural Sciences and Engineering Research Council of Canada (#6793); KBS holds the Canada Research Chair in Molecular Physiology. The authors thank J.M. Storey for editorial review of manuscript, and R. Al-Attar for providing technical support.

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  1. Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, ON, K1S 5B6, Canada

    Gurjit Singh & Kenneth B. Storey

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Singh, G., Storey, K.B. MondoA:MLX complex regulates glucose-dependent gene expression and links to circadian rhythm in liver and brain of the freeze-tolerant wood frog, Rana sylvatica. Mol Cell Biochem 473, 203–216 (2020). https://doi.org/10.1007/s11010-020-03820-9

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