Cytochrome P450 diversity and induction by gorgonian allelochemicals in the marine gastropod Cyphoma gibbosum
Additional file 1: Summary of digestive gland RNA and protein samples collected during January 2006 feeding assays. (71.67Kb)
Additional file 3: Degenerate and specific oligonucleotide primers for initial and full-length amplification of Cyphoma gibbosum CYP4s and actin. (71.62Kb)
Additional file 6: Maximum parsimony trees depicting transcripts recognized by CYP4BL(subA) and CYP4BL(subB) quantitative RT-PCR primers. (181.9Kb)
Additional file 8: Oligonucleotide primers for recombinant C. gibbosum CYP4 expression in yeast. (65.29Kb)
Additional file 12: Results of a one-way MANOVA investigating CYP4 gene expression variability among reefs for time-zero and control-fed C. gibbosum. (87.19Kb)
Additional file 13: Results of a two-way MANOVA investigating differences in digestive gland CYP4 gene expression in C. gibbosum feeding on control versus gorgonian diets. (93.42Kb)
Additional file 14: Results of ANOVA comparisons (Univariate F-tests) of diet- and reef-specific mean CYP4 gene expression in C. gibbosum feeding on control vs. gorgonian diets. (100.6Kb)
Additional file 16: Leukotriene B4 hydroxylase activity of heterologously expressed Cyphoma CYP4 proteins. (67.89Kb)
Additional file 17: Homology models CYP4BL3 and CYP4BK1 depicting putative substrate access channels. (527.7Kb)
Whalen, Kristen E.
Starczak, Victoria R.
Nelson, David R.
Goldstone, Jared V.
Hahn, Mark E.
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Intense consumer pressure strongly affects the structural organization and function of marine ecosystems, while also having a profound effect on the phenotype of both predator and prey. Allelochemicals produced by prey often render their tissues unpalatable or toxic to a majority of potential consumers, yet some marine consumers have evolved resistance to host chemical defenses. A key challenge facing marine ecologists seeking to explain the vast differences in consumer tolerance of dietary allelochemicals is understanding the biochemical and molecular mechanisms underlying diet choice. The ability of marine consumers to tolerate toxin-laden prey may involve the cooperative action of biotransformation enzymes, including the inducible cytochrome P450s (CYPs), which have received little attention in marine invertebrates despite the importance of allelochemicals in their evolution. Here, we investigated the diversity, transcriptional response, and enzymatic activity of CYPs possibly involved in allelochemical detoxification in the generalist gastropod Cyphoma gibbosum, which feeds exclusively on chemically defended gorgonians. Twelve new genes in CYP family 4 were identified from the digestive gland of C. gibbosum. Laboratory-based feeding studies demonstrated a 2.7- to 5.1-fold induction of Cyphoma CYP4BK and CYP4BL transcripts following dietary exposure to the gorgonian Plexaura homomalla, which contains high concentrations of anti-predatory prostaglandins. Phylogenetic analysis revealed that C. gibbosum CYP4BK and CYP4BL were most closely related to vertebrate CYP4A and CYP4F, which metabolize pathophysiologically important fatty acids, including prostaglandins. Experiments involving heterologous expression of selected allelochemically-responsive C. gibbosum CYP4s indicated a possible role of one or more CYP4BL forms in eicosanoid metabolism. Sequence analysis further demonstrated that Cyphoma CYP4BK/4BL and vertebrate CYP4A/4F forms share identical amino acid residues at key positions within fatty acid substrate recognition sites. These results demonstrate differential regulation of CYP transcripts in a marine consumer feeding on an allelochemical-rich diet, and significantly advance our understanding of both the adaptive molecular mechanisms that marine consumers use to cope with environmental chemical pressures and the evolutionary history of allelochemical-metabolizing enzymes in the CYP superfamily.
© The Authors, 2010. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in BMC Ecology 10 (2010): 24, doi:10.1186/1472-6785-10-24.
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