Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Molecular Characterization, Protein-protein Interaction Network, and Evolution of four Glutathione Peroxidases from Tetrahymena Thermophila

Version 1 : Received: 7 September 2020 / Approved: 8 September 2020 / Online: 8 September 2020 (04:59:18 CEST)

How to cite: Ferro, D.; Bakiu, R.; Pucciarelli, S.; Miceli, C.; Vallesi, A.; Irato, P.; Santovito, G. Molecular Characterization, Protein-protein Interaction Network, and Evolution of four Glutathione Peroxidases from Tetrahymena Thermophila. Preprints 2020, 2020090171 (doi: 10.20944/preprints202009.0171.v1). Ferro, D.; Bakiu, R.; Pucciarelli, S.; Miceli, C.; Vallesi, A.; Irato, P.; Santovito, G. Molecular Characterization, Protein-protein Interaction Network, and Evolution of four Glutathione Peroxidases from Tetrahymena Thermophila. Preprints 2020, 2020090171 (doi: 10.20944/preprints202009.0171.v1).

Abstract

Glutathione peroxidases (GPxs) form a broad family of antioxidant proteins essential for maintaining redox homeostasis in eukaryotic cells. In this study, we used an integrative approach that combines bioinformatics, molecular biology, and biochemistry to investigate the role of GPxs in reactive oxygen species detoxification in the unicellular eukaryotic model organism Tetrahymena thermophila. Both phylogenetic and mechanistic empirical model analyses provided indications about the evolutionary relationships among the GPXs of Tetrahymena and the orthologous enzymes of phylogenetically related species. In-silico gene characterization and text mining were used to predict the functional relationships between GPxs and other physiologically-relevant processes. The GPx genes contain conserved transcriptional regulatory elements in the promoter region, which suggest that transcription is under tight control of specialized signaling pathways. The bioinformatic findings were next experimentally validated by studying the time course of copper (Cu)-dependent regulation of gene transcription and enzymatic activity. Results emphasize the role of GPxs in the detoxification pathways that, by complex regulation of Cu-dependent GPx gene expression, enables Tetrahymena to survive in high Cu concentrations and the associated redox environment.

Subject Areas

protein-protein interaction network; GPx; glutathione peroxidases genes; ciliate protists; copper; metals; antioxidant system; free-radicals; ROS; reactive oxygen species

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