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Molecular Characterization of Metallothionein Gene of the Korean Bitterling Acheilognathus signifer (Cyprinidae)  

Lee, Sang-Yoon (Department of Marine Bio-Materials & Aquaculture, Pukyong National University)
Bang, In-Chul (Department of Marine Biotechnology, Soonchunhyang University)
Nam, Yoon-Kwon (Department of Marine Bio-Materials & Aquaculture, Pukyong National University)
Publication Information
Korean Journal of Ichthyology / v.23, no.1, 2011 , pp. 10-20 More about this Journal
Abstract
Genetic determinant for metallothionein (MT), a cysteine-rich protein playing essential roles in metal detoxification and homeostasis, was characterized in the Korean bitterling (Acheilognathus signifer, Cyprinidae), an endemic fish species. The full-length A. signifer MT (AsMT) cDNA (551 bp) is composed of a single open-reading frame (ORF) to encode a polypeptide of 60 amino acids containing 20 cysteine residues whose positions are conserved in most cypriniform MTs. At the genomic level, the AsMT (2,593 bp spanning the 5'-flanking region to the 3'-untranslated region) represented a conserved tripartite (three exons interrupted by two introns) structure with AT-rich introns. The upstream regulatory region (-1,914 bp from the ATG initiation codon) of AsMT displayed various sites and motifs for transcription factors involved in the metal-mediated regulation and stress/immune responses. The AsMT transcript was ubiquitously detected in various organs with variable expression levels, where the ovary and intestine showed the highest expression, while the heart and skeletal muscle represented the lowest level. During an exposure to copper (immersion in $0.5\;{\mu}M$ Cu for 48 h), the levels of AsMT transcripts were significantly elevated in the liver (more than 3.5-fold), moderately in the gill, kidney, and spleen (ranging from 1.5- to 2.5-fold), and barely in the brain and intestine. Results of this study could form a useful basis to explore the metal-related stress physiology of this endangered fish species.
Keywords
Gene structure and expression; Korean bitterling; Acheilognathus signifer; metallothionein;
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