Abstract
Knowledge on antifungal biomolecules is limited compared to antibacterial peptides. A strictly antifungal peptide from the blue mussel, Mytilus edulis named mytimycin (MytM) was reported in 1996 as partial NH2 33 amino acid sequence. Using back-translations of the previous sequence, MytM-related nucleotide sequences were identified from a normalized Mytilus galloprovincialis expressed sequence tag library. Primers designed from a consensus sequence have been used to obtain a fragment of 560 nucleotides, including the complete coding sequence of 456 nucleotides. Precursor is constituted by a signal peptide of 23 amino acids, followed by MytM of 54 amino acids (6.2–6.3 kDa, 12 cysteines) and C-terminal extension of 75 amino acids. Only two major amino acid precursor sequences emerged, one shared by M. galloprovincialis from Venice and Vigo, the other belonging to M. galloprovincialis from Palavas, with nine amino acid differences between the two MytM. Predicted disulfide bonds suggested the presence of two constrained domains joined by amino acidic NIFG track. Intriguing was the presence of conserved canonical EF hand-motif located in the C-terminus extension of the precursor. The MytM gene was found interrupted by two introns. Intron 2 existed in two forms, a long (1,112 nucleotides) and a short (716 nucleotides) one resulting from the removal of the central part of the long one. Both the short (GenBank FJ804479) and the long (GenBank FJ804478) genes are simultaneously present in the mussel genome.
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Abbreviations
- EST:
-
Expressed sequence tag
- gDNA:
-
Genomic DNA
- MytM:
-
Mature moiety of mytimycin precursor
- MytM-P:
-
Mature moiety of mytimycin from Palavas
- MytM-V:
-
Mature moiety of mytimycin from Vigo and Venice
- UTR:
-
Untranslated region
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Acknowledgments
This collaborative work was partly funded by the EU program IMAQUANIM (FOOD-CT-2005-007103). MT was fully supported by IMAQUANIM. Thanks to Umberto Rosani for the preliminary comparison performed on new FLX 454 sequence MytM data.
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Table 1
Supplementary data. Nucleotides changes within mytimycin precursor observed in M. galloprovincialis from Palavas, France; Venice, Italy; and Vigo, Spain with the involved codons, the number of clones and corresponding amino acid. MytM is gray boxed. Changed nucleotides are in bold–gray boxed. Amino acids in italic referred to equivalent replacements in terms of side chain characteristics. Note the high number of changes in Vigo’s mussels, eight of them being silent and eight other corresponding to replacements by equivalent amino acids. Note also that the only four changes in Palavas’ mussels were common to Vigo’s mussels, whereas only one change out of five from Venice’s mussels was shared with Vigo’s mussels. Except for locations +299/+300, note that one codon was always largely predominant (DOC 123 kb)
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Sonthi, M., Toubiana, M., Pallavicini, A. et al. Diversity of Coding Sequences and Gene Structures of the Antifungal Peptide Mytimycin (MytM) from the Mediterranean Mussel, Mytilus galloprovincialis . Mar Biotechnol 13, 857–867 (2011). https://doi.org/10.1007/s10126-010-9345-4
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DOI: https://doi.org/10.1007/s10126-010-9345-4