Abstract
Pyrococcus furiosus is a remarkable archaeon able to grow at temperatures around 100 °C. To gain insight into how this model hyperthermophile copes with heat stress, we compared transcriptomic and metabolomic data of cells subjected to a temperature shift from 90 °C to 97 °C. In this study, we used RNA-sequencing to characterize the global variation in gene expression levels, while nuclear magnetic resonance (NMR) and targeted ion exchange liquid chromatography–mass spectrometry (LC–MS) were used to determine changes in metabolite levels. Of the 552 differentially expressed genes in response to heat shock conditions, 257 were upregulated and 295 were downregulated. In particular, there was a significant downregulation of genes for synthesis and transport of amino acids. At the metabolite level, 37 compounds were quantified. The level of di-myo-inositol phosphate, a canonical heat stress solute among marine hyperthermophiles, increased considerably (5.4-fold) at elevated temperature. Also, the levels of mannosylglycerate, UDP-N-acetylglucosamine (UDPGlcNac) and UDP-N-acetylgalactosamine were enhanced. The increase in the pool of UDPGlcNac was concurrent with an increase in the transcript levels of the respective biosynthetic genes. This work provides the first metabolomic analysis of the heat shock response of a hyperthermophile.
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Acknowledgements
This work was supported by: Project LISBOA-01-0145-FEDER-007,660 (Microbiologia Molecular, Estrutural e Celular) funded by FEDER through COMPETE2020—Programa Operacional Competitividade e Internacionalização (POCI) and by national funds from FCT—Fundação para a Ciência e a Tecnologia and by project ONEIDA (LISBOA-01-0145-FEDER-016,417) co-funded by FEEI—“Fundos Europeus Estruturais e de Investimento” from “Programa Operacional Regional Lisboa 2020” and by national funds from FCT. The NMR equipment at CERMAX is part of the National NMR Network and is partially supported by Infrastructure Project no. 022,161 (co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC). Mass Spectrometry investigations were carried out at MetaToul (Metabolomics & Fluxomics Facitilies, Toulouse, France, www.metatoul.fr) which is part of MetaboHUB (The French National infrastructure for metabolomics and fluxomics, www.metabohub.fr, MetaboHUB-ANR-11-INBS-0010). MetaToul is supported by grants from the Région Occitanie, the FEDER (through FEDER-FSE 2014-2020), Toulouse Metropole, the Infrastructures en Biologie, Santé et Agronomie (IBiSa, France), the Centre National de la Recherche Scientifique (CNRS) and the Institut National de la Recherche Agronomique (INRA). We thank D. L. Turner for critical reading of the manuscript.
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Esteves, A.M., Graça, G., Peyriga, L. et al. Combined transcriptomics–metabolomics profiling of the heat shock response in the hyperthermophilic archaeon Pyrococcus furiosus. Extremophiles 23, 101–118 (2019). https://doi.org/10.1007/s00792-018-1065-0
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DOI: https://doi.org/10.1007/s00792-018-1065-0