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Molecular cloning and characterization of calreticulin from rainbow trout (Oncorhynchus mykiss)

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Abstract

Calreticulin (CRT) is a highly conserved, high-capacity, calcium-binding protein shared among vertebrates, invertebrates and higher plants. Its biological importance, highlighted by its highly conserved nature, is supported by its crucial physiological and immunological functions. Within the endoplasmic reticulum, CRT serves as a calcium modulator and a lectin-like chaperone for glycoproteins, especially class I major histocompatibility receptors. To date, CRT cDNA clones have been isolated from a wide range of phyla, yet little is known about this gene in fish species, the largest and most diverse group of jawed vertebrates. This report describes the cloning of a cDNA from a rainbow trout pronephros library that encodes a deduced 419-amino acid protein, which includes a predicted 20-amino acid signal peptide and has a 69% amino acid identity to both murine and human CRT. Like its mammalian counterparts, this cDNA contains conserved cysteine residues believed to form a disulphide bond, a proline-rich region which includes a potential N-glycosylation site, and a highly acidic C-terminal domain terminating with the endoplasmic reticulum retrieval sequence, KDEL. Reverse transcription tissue-distribution assays indicate it is ubiquitously expressed in all tissues tested with highest expression in liver, while Southern blotting indicates it is a single copy gene.

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Acknowledgements

The authors would like to thank Dr. Tom McConnell for kindly sharing sequence data prior to publication and Dr. Roy Sundick for providing the rainbow trout cDNA library. This work was supported in part by grant number CRD226816-99 to B.D. from the Natural Sciences and Engineering Research Council of Canada, AquaBounty Canada, Inc. and the AquaNet Network Centre of Excellence.

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Authors and Affiliations

  1. Department of Biology, University of Waterloo, 200 University Avenue, West, Waterloo, ON, N2L 3G1, Canada

    Stephen Kales, Kazuhiro Fujiki & Brian Dixon

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  1. Stephen Kales
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  2. Kazuhiro Fujiki
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  3. Brian Dixon
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Corresponding author

Correspondence to Brian Dixon.

Additional information

The nucleotide sequences reported here have been submitted to the GenBank database with the following accession numbers: AY372389 and AY372527. The accession numbers of CRT sequences retrieved in this study are as follow: Atlantic salmon, EST-BI468042; channel catfish, AY342298 (sequence courtesy of T. McConnell, East Carolina University); zebrafish, AF195882; zebrafish MGC:55972 clone, BC046906; lamprey fragment, AB025328; hagfish fragment, AB025323; mouse, BC003453; C. elegans, NM_072174; corn, AF190454: Arabidopsis, NM_104513; Dictyostelium, U36937; human, BT007448; and Xenopus, BC046699.

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Kales, S., Fujiki, K. & Dixon, B. Molecular cloning and characterization of calreticulin from rainbow trout (Oncorhynchus mykiss). Immunogenetics 55, 717–723 (2004). https://doi.org/10.1007/s00251-003-0631-4

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  • DOI: https://doi.org/10.1007/s00251-003-0631-4

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