Molecular Characterization, Expression Profiles, and Immunostimulation Responses of TRAF6 and TAK1 in Japanese Flounder (Paralichthys olivaceus)
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Tumor necrosis factor receptor-associated factor 6 (TRAF6) and transforming growth factor-β-activated kinase 1 (TAK1) are two important adaptor molecules in Toll-like receptor (TLR) signaling pathway. In this study, TRAF6 (PoTRAF6) and TAK1 (PoTAK1) were cloned and characterized in Japanese flounder (Paralichthys olivaceus). The full-length cDNA sequence of PoTRAF6 is 1953 bp, with an open reading frame (ORF) of 1713 bp encoding a putative protein of 570 amino acids. PoTRAF6 contains one really interesting new gene (RING) domain, two zinc fingers, one coiled-coil region, and one meprin and TRAF homology (MATH) domain, which shows a high similarity to TRAF6s in other species. The full-length PoTAK1 cDNA sequence is 2086 bp, with an ORF of 1728 bp that encodes a putative protein of 575 amino acids. PoTAK1 contains a conserved serine/threonine protein kinase catalytic domain and a coiled-coil region. The promoter regions of PoTRAF6 and PoTAK1 were also analyzed to predict several potential transcription factor-binding sites. In addition, the expression patterns of these two genes were examined in developmental stages, different tissues, and challenged samples. PoTRAF6 and PoTAK1 were expressed during the whole developmental stages, and the highest expressions were in intestine and heart, respectively. In challenged embryonic cells with LPS, CpG ODN, and poly I:C, the expressions of PoTRAF6 and PoTAK1 were both up-regulated significantly. These results suggest that PoTRAF6 and PoTAK1 play crucial roles in immune responses and may be involved in the developmental process of Japanese flounder.
Key wordsTRAF6 TAK1 Paralichthys olivaceus cloning expression promoter analysis immunostimulation
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This study was supported by the National Natural Science Foundation of China (No. 31101891) and the National High-tech Research and Development Program (No. 2012AA10A402).
- Angel, P., Imagawa, M., Chiu, R., Stein, B., Imbra, R. J., Rahmsdorf, H. J., Jonat, C., Herrlich, P., and Karin, M., 1987. Phorbol ester–inducible genes contain a common cis element recognized by a TPA–modulated trans–acting factor. Cell Press, 49 (6): 729–739.Google Scholar
- Basu, M., Swain, B., Maiti, N. K., Routray, P., and Samanta, M., 2012. Inductive expression of toll–like receptor 5 (TLR5) and associated downstream signaling molecules following ligand exposure and bacterial infection in the Indian major carp, mrigal (Cirrhinus mrigala). Fish & Shellfish Immunology, 32 (1): 121–131.CrossRefGoogle Scholar
- Chung, J. Y., Park, Y. C., Ye, H., and Wu, H., 2002. All TRAFs are not created equal: common and distinct molecular mechanisms of TRAF–mediated signal transduction. Journal of Cell Science, 115 (4): 679–688.Google Scholar
- Delaney, J. R., and Mlodzik, M., 2006. TGF–beta activated kinase–1. New insights into the diverse roles of TAK1 in development and immunity. Cell Cycle, 5 (24): 2852–2855.Google Scholar
- Felix, J., and Savvides, S. N., 2017. Mechanisms of immunomodulation by mammalian and viral decoy receptors: Insights from structures. Nature Reviews: Immunology, 17 (2): 112–129.Google Scholar
- Ha, H., Han, D., and Choi, Y., 2009. TRAF–mediated TNFRfamily signaling. Current Protocols in Immunology, 11: 9.Google Scholar
- Medzhitov, R., 2001. Toll–like receptors and innate immunity. Nature Reviews: Immunology, 1 (2): 135.Google Scholar
- Mizushima, S., Ishida, T., Azuma, S., Kobayashi, N., Tojo, T., Suzuki, K., Aizawa, S., Watanabe, T., Mosialos, G., Kieff, E., Yamamoto, T., and Inoue, Ji., 1996. Identification of TRAF6, a novel tumor necrosis factor receptor–associated factor protein that mediates signaling from an amino–terminal domain of the CD40 cytoplasmic region. Journal of Biological Chemistry, 271 (46): 28745–28748.CrossRefGoogle Scholar
- Nho, S. W., Hikima, J., Cha, I. S., Park, S. B., Jang, H. B., Castillo, C. S., Kondo, H., Hirono, I., Aoki, T., and Jung, T. S., 2011. Complete genome sequence and immunoproteomic analyses of the bacterial fish pathogen Streptococcus parauberis. Journal of Bacteriology, 193: 3356–3366.CrossRefGoogle Scholar
- Tanekhy, M., Matsuda, S., Itano, T., Kawakami, H., Kono, T., and Sakai, M., 2010. Expression of cytokine genes in head kidney and spleen cells of Japanese flounder (Paralichthys olivaceus) infected with Nocardia seriolae. Veterinary Immunology and Immunopathology, 134 (3–4): 178–183.CrossRefGoogle Scholar
- Ye, H., Arron, J. R., Lamothe, B., Cirilli, M., Kobayashi, T., Shevde, N. K., Segal, D., Dzivenu, O. K., Vologodskaia, M., Yim, M., Du, K., Singh, S., Pike, J. W., Darnay, B. G., Choi, Y., and Wu, H., 2002. Distinct molecular mechanism for initiating TRAF6 signalling. Nature, 418 (6896): 443–447.CrossRefGoogle Scholar