Isolation and characterization of two genes encoding calitoxins, neurotoxic peptides from Calliactis parasitica (Cnidaria).
Among sea anemone neurotoxins, calitoxin, recently isolated from Calliactis parasitica, is a highly toxic peptide of 46 amino acids (aa), whose sequence differs greatly from that of all sea anemone toxins isolated so far. In this study, two genes (clx-1 and clx-2) coding for two highly homologous calitoxins were isolated and characterized from a C. parasitica genomic library. The clx-1 gene encodes the already known calitoxin sequence, named CLX-I, whereas a single bp substitution in the coding region of clx-2 is responsible for a single Glu6-->Lys replacement in a new peptide named CLX-II. The structural organization of the two genes is very similar: two introns and three exons, whose sequences are highly homologous for clx-1 and clx-2 (95% identity). The open reading frame (ORF) of both clx-1 and clx-2 codes for a precursor peptide of 79 aa, whose N-terminus has the feature of a single peptide, while the C-terminus corresponds to the sequences of mature CLX-I and CLX-II. The finding that a pair of basic aa is located upstream from the sequence of both mature toxins strongly suggests that proteolytic events, at specific cleavage sites, are responsible for the release of neurotoxins from their respective precursor molecules.
Spagnuolo
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Molecular Biology and Biochemistry Laboratory, Stazione Zoologica, A. Dohrn, Naples, Italy.
Zanetti
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Cariello
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Piccoli
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003064:Cnidarian Venoms / Q000096:biosynthesis / Q000235:genetics*; D004247:DNA / Q000235:genetics; D017931:DNA Primers; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000096:biosynthesis; D011061:Poly A / Q000302:isolation & purification / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D012333:RNA, Messenger; D015183:Restriction Mapping; D012615:Sea Anemones / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D003064:Cnidarian Venoms; D017931:DNA Primers; D009498:Neurotoxins; D012333:RNA, Messenger; C060036:calitoxin; D011061:Poly A; D012313:RNA; D004247:DNA