Characterization of genes involved in ceramide metabolism in the Pacific oyster (Crassostrea gigas)

BMC Research Notes

Steven Roberts¹  Emma Timmins-Schiffman¹ 

[1] School of Aquatic and Fishery Sciences, University of Washington, Seattle, WA, 98105, USA

关键词: Bacteria;    Ceramide;    Stress;    Crassostrea gigas;    Oyster;   

Others  :  1165712
DOI  :  10.1186/1756-0500-5-502

 received in 2012-06-01, accepted in 2012-09-06,  发布年份 2012

【 摘 要 】

Background

The lipid signaling molecule, ceramide, is a key component of the vertebrate stress response, however, there is limited information concerning its role in invertebrate species. In order to identify genes involved in ceramide metabolism in bivalve molluscs, Pacific oyster genomic resources were examined for genes associated with ceramide metabolism and signaling.

Results

Several genes were identified including full-length sequences characterized for serine palmitoyltransferase-1, 3-ketodihydrosphingosine reductase, acid ceramidase,and ceramide glucosyltransferase. Genes involved in ceramide synthesis and metabolism are conserved across taxa in both form and function. Expression analysis as assessed by quantitative PCR indicated all genes were expressed at high levels in gill tissue. The role of the ceramide pathway genes in the invertebrate stress response was also explored by measuring expression levels in adult oysters exposed to Vibrio vulnificus. Two genes demonstrated increased expression during the bacterial challenge: a gene involved in hydrolytic breakdown of ceramide (acid ceramidase) and a gene involved in de novo generation of ceramide (3-ketodihydrosphingosine reductase), suggesting a possible role of ceramide in the invertebrate stress and immune responses.

Conclusions

In silico and laboratory results support that Pacific oysters have the basic components of the ceramide metabolism pathway. These results also indicate that ceramide may have analogous functions in vertebrates and invertebrates. The gene expression pattern of acid ceramidase and 3-kethodihydrosphingosine reductase in response to bacterial exposure especially supports that ceramide and sphingolipid metabolism may be involved in the oyster’s stress and/or immune responses.

【 授权许可】

   
2012 Timmins-Schiffman and Roberts; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Hannun YA: The sphingomyelin cycle and the second messenger function of ceramide. J Biochem Chem 1994, 269:3125-3128.
• [2]Ballou LR, Laulederkind SJ, Rosloniec EF, Raghow R: Ceramide signaling and the immune response. Biochim Biophys Acta 1996, 1301:273-287.
• [3]Eliyahu E, Park J-H, Shtraizent N, He X, Schuchman EH: Acid ceramidase is a novel factor required for early embryo survival. FASEB J 2007, 21:1403-1409.
• [4]Chavez JA, Holland WL, Bär J, Sandhoff K, Summers SA: Acid ceramidase overexpression prevents the inhibitory effects of saturated fatty acids on insulin signaling. J Biol Chem 2005, 280:20148-20153.
• [5]Perry DK, Bielawska A, Hannun YA: Quantitative determination of ceramide using diglyceride kinase. Methods Enzymol 2000, 312:22-31.
• [6]Strelow A, Bernardo K, Adam-Klages S, Linke T, Sandhoff K, Krönke M, Adam D: Overexpression of acid ceramidase protects from tumor necrosis factor-induced cell death. J Exp Med 2000, 192:601-612.
• [7]El Babili M, Brichon G, Zwingelstein G: Sphingomyelin metabolism is linked to salt transport in the gills of euryhaline fish. Chem Mater Sci 1996, 31:385-392.
• [8]Verheij M, Bose R, Lin XH, Yao B, Jarvis WD, Grant S, Birrer MF, Szabo E, Zon LI, Kyriakis JM, Haimovitz-Friedman A, Fuks Z, Kolesnick RN: Requirement for ceramide-initiated SAPK/JNK signaling in stress-induced apoptosis. Lett Nat 1996, 380:75-79.
• [9]Ballou LR, Chao CP, Holness MA, Barker SC, Raghow R: Interleukin-1-mediated PGE2 production and sphingomyelin metabolism. Evidence for the regulation of cyclooxygenase gene expression by sphingosine and ceramide. J Biol Chem 1992, 267:20044-20050.
• [10]Zhao S, Fernald RD: Comprehensive algorithm for quantitative real-time polymerase chain reaction. Comput Biol 2005, 12:1045-1062.
• [11]Kihara A, Igarashi Y: FVT-1 is a mammalian 3-Ketodihydrosphingosine Reductase with an active site that faces the cytosolic side of the endoplasmic reticulum membrane. J Biol Chem 2004, 279:49243-49250.
• [12]Lee Y, De Zoysa M, Whang I, Lee S, Kim Y, Oh C, Choi CY, S-Y Y, Lee J: Molluscan death effector domain (DED)-containing caspase-8 gene from disk abalone (Haliotis discus discus): Molecular characterization and expression analysis. Fish Shellfish Immunol 2011, 30:480-487.
• [13]Zhang L, Li L, Zhang G: Gene discovery, comparative analysis and expression profile reveal the complexity of the Crassostrea gigas apoptosis system. Dev Comp Immunol 2011, 35:603-610.
• [14]Romero A, Estévez-Calvar N, Dios S, Figueras A, Novoa B: New insights into the apoptotic process in mollusks: characterization of caspase genes in Mytilus galloprovincialis. PLoS One 2011, 6:e17003.
• [15]Hannun YA, Luberto C: Ceramide in the eukaryotic stress response. Trends Cell Biol 2000, 10:73-80.
• [16]Perry DK: Serine palmitoyltransferase: role in apoptotic de novo ceramide synthesis and other stress responses. Biochimica et Biophysica Acta – Molecular and Cell Biology of. Lipids 2002, 1585:146-152.
• [17]Hanada K: Serine palmitoyltransferase, a key enzyme of sphingolipid metabolism. Biochimica et Biophysica Acta – Molecular and Cell Biology of. Lipids 2003, 1632:16-30.
• [18]Momany C, Ghosh R, Hackert ML: Structural motifs for pyridoxal-5’-phosphate binding in decarboxylases: An analysis based on the crystal structure of the Lactobacillus 30a ornithine decarboxylase. Protein Sci 2008, 4:849-854.
• [19]Merrill AH Jr, Stevens VL: Modulation of protein kinase C and diverse cell functions by sphingosine – a pharmacologically interesting compound linking sphingolipids and signal transduction. Biochim Biophys Acta 1989, 1010:131-139.
• [20]Baumruker T, Prieschl EE: Sphingolipids and the regulation of the immune response. Semin Immunol 2002, 14:57-63.
• [21]Philipp EER, Kraemer L, Melzner F, Poustka AJ, Thieme S, Findeisen U, Schreiber S, Rosenstiel P: Massively Parallel RNA Sequencing Identifies a Complex Immune Gene Repertoire in the lophotrocozoan Mytilus edulis. PLoS One 2012, 7:e33091.
• [22]Lackner DH, Schmidt MW, Wu S, Wolfe DA, Bahler J: Regulation of transcriptome, translation, and proteome in response to environmental stress in fission yeast. Genome Biol 2012, 13:R25. BioMed Central Full Text
• [23]Le Grand F, Kraffe E, Marty Y, Donaghy L, Soudant P: Membrane phospholipid composition of hemocytes in the Pacific oyster Crassostrea gigas and the Manila clam Ruditapes philippinarum. Comparative Biochemistry and Physiology – Part A: Molecular and Integrative. Physiology 2011, 159:383-391.
• [24]Altschul SF, Madden TL, Schaeffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ: Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 1997, 25:3389-3402.
• [25]Drummond AJ, Ashton B, Buxton S, Cheung M, Cooper A, Heled J, Kearse M, Moire R, Stones-Havas S, Sturrock S, Thierer T, Wilson A: Geneious v4.5.6. 2010. Available from http://www.geneious.com webcite
• [26]Rozen S, Skaletsky HJ: Primer3 on the WWW for general users and for biologist programmers. In Bioinformatics Methods and Protocols: Methods in Molecular Biology. Edited by Drawetz S, Misener S. Humana Press, Totowa, NJ; 2000:365-386.
• [27]Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DF: Clustal W and Clustal X version 2.0. Bioinformatics 2007, 23:2947-2948.
• [28]Guindon S, Gascuel O: A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 2003, 52:696-704.
• [29]Jones DT, Taylor WR, Thornton JM: The rapid generation of mutation data matrices from protein sequences. Comput Appl Biosci 1992, 8:275-282.
• [30]R Development Core Team: R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria; 2011. www.R-project.org webcite