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Comparative molecular characterization of the regucalcin (RGN) gene in rainbow trout (Oncorhynchus mykiss) and maraena whitefish (Coregonus marena)

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Abstract

The Ca2+-binding protein regucalcin (RGN) is crucial for the regulation of Ca2+ ion homeostasis and signal transduction of cells. It is involved in the regulation of Ca2+-dependent protein kinases and Ca2+ pump enzymes in cell membranes. Comparative transcriptome analysis in healthy fish of two aquacultured rainbow trout (Oncorhynchus mykiss) lines (BORN, TCO) varying in susceptibility to environmental stress identified significant differences in the expression of the RGN gene. Therefore, we firstly determined the full genomic DNA and cDNA sequence of RGN gene from rainbow trout and comparatively investigated the complete cDNA sequence in another salmonid fish dedicated for local aquaculture, the maraena whitefish (Coregonus marena). The sequence coding region translates for proteins of 298 and 299 amino acids (aa), respectively, indicating a high conservation of RGN proteins (95.7% aa identity) between the two related salmonids. In the second place, we generated RGN gene expression profiles after pathogen (Aeromonas salmonicidae subsp. salmonicida) and temperature (8 and 23°C) challenge in the two rainbow trout lines using salmon microarrays and quantitative RT-PCR. The profiles not only verified initially detected gene expression differences, they also display a tissue specific gene expression in dependence from the stressor and time. The differences in gene expression support our assumption that RGN might play a role in recovery of rainbow trout after environmental stress.

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Acknowledgments

We are grateful to B. Schöpel, I. Hennings, and M. Fuchs for excellent technical assistance. This work was funded by the Exzellenzförderprogramm Mecklenburg-Vorpommern 2008–2010 (project AU08026 entitled DIREFO).

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Authors and Affiliations

  1. Leibniz-Institut für Nutztierbiologie (FBN), Fachbereich Molekularbiologie, Wilhelm-Stahl-Allee 2, 18196, Dummerstorf, Germany

    Marieke Verleih, Alexander Rebl, Klaus Wimmers & Tom Goldammer

  2. Friedrich-Loeffler-Institut (FLI), Institut für Infektionsmedizin, Greifswald, Insel Riems, Germany

    Bernd Köllner & Tomáš Korytář

  3. Landesforschungsanstalt für Landwirtschaft und Fischerei Mecklenburg-Vorpommern (LFA-MV), Institut für Fischerei, Born, Germany

    Eckhard Anders

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  1. Marieke Verleih
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  2. Alexander Rebl
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  3. Bernd Köllner
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  4. Tomáš Korytář
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  5. Eckhard Anders
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  6. Klaus Wimmers
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  7. Tom Goldammer
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Correspondence to Tom Goldammer.

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11033_2011_1216_MOESM1_ESM.tif

Supplementary Figure S1 Alignment of predicted RGN protein sequences from rainbow trout and maraena whitefish with all known teleostean homologues as well as respective sequences of higher vertebrates. Accession numbers of used proteins and abbreviations are listed in Table 2. Identical and strongly similar amino acids are labeled by black and dark gray underlay as well as white letters; similar amino acids are marked by light gray underlay. Conceptionally translated sequences, which are derived in this study, are written in bold letters. The localization of the SGL multi-domain (SMP-30/Gluconolaconase/LRE-like region) is marked by a black bracket. Black arrows indicate conserved positions of Ca2+ coordination sides at position E18, N154 and D204. (TIFF 7100 kb)

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Verleih, M., Rebl, A., Köllner, B. et al. Comparative molecular characterization of the regucalcin (RGN) gene in rainbow trout (Oncorhynchus mykiss) and maraena whitefish (Coregonus marena). Mol Biol Rep 39, 4291–4300 (2012). https://doi.org/10.1007/s11033-011-1216-1

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  • DOI: https://doi.org/10.1007/s11033-011-1216-1

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