title: Molecular cloning, expression and functional characterization of tumor necrosis factor (TNF) receptor-associated factor (TRAF)-interacting protein (TRIP) in grass carp, Ctenopharyngodon idella.
abstract: TRIP (Tumor Necrosis Factor (TNF) Receptor-Associated Factor (TRAF)-Interacting Protein), a member of the TNF superfamily, plays a crucial role in the modulation of inflammation in vertebrates. However, no information about TRIP is available in teleosts. In this study, the full-length cDNA of TRIP, containing a 5'UTR of 112 bp, an ORF of 1359 bp, and a 3'UTR of 29 bp before the poly (A) tail, was cloned from grass carp, Ctenopharyngodon idella. The TRIP gene encoded a protein of 452 amino acids with an estimated molecular mass of 51.06 KD and a predicted theoretical isoelectric point (pI) of 9.11. Quantitative real-time PCR analysis revealed that TRIP mRNA was expressed in all the tissues examined in grass carp, with the highest expression in the kidney, followed by the intestine and thymus. However, lower levels of expression were also detected in fat, spleen, liver, gonad and heart. Subcellular localization and two-hybrid analysis revealed that TRIP was located in the nucleus and that it interacted with TRAF1 and TRAF2 in HEK293T cells. Furthermore, similar to TNF-α, IL-10 and TRIP mRNA expression was upregulated in the spleen of fish fed high-fat or high-carbohydrate diets, suggesting that TRIP might be associated with the response to excessive energy intake. The mRNA relative expression of TRIP was significantly reduced (P < 0.05) after hepatocyte of C. idella was treated with 2 μg/mL lipopolysaccharide (LPS) for 4 h, while the expression levels of inflammatory cytokines TNF-α and IL-10 were significantly increased (P < 0.05). Taken together, these results indicate that TRIP might play important roles in immune defense and has the potential to be used as a anti-inflammation target in grass carp.
authors.affiliation: College of Fisheries, Henan Normal University, Xinxiang 453007, China.
authors.forename: R-H
authors.initials: RH
authors.lastname: Lu
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.
authors.forename: Z-G
authors.initials: ZG
authors.lastname: Chang
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China.
authors.forename: J
authors.initials: J
authors.lastname: Sun
authors.affiliation: College of Fisheries, Henan Normal University, Xinxiang 453007, China.
authors.forename: F
authors.initials: F
authors.lastname: Yang
authors.affiliation: College of Fisheries, Henan Normal University, Xinxiang 453007, China.
authors.forename: G-X
authors.initials: GX
authors.lastname: Nie
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China. Electronic address: jihong@nwsuaf.edu.cn.
authors.forename: H
authors.initials: H
authors.lastname: Ji
mesh_terms: D000821:Animal Feed / Q000032:analysis; D000818:Animals; D002347:Carps / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004032:Diet / Q000662:veterinary; D004040:Dietary Carbohydrates / Q000008:administration & dosage*; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D008055:Lipids / Q000008:administration & dosage*; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020539:Sequence Analysis, Protein / Q000662:veterinary; D047988:Tumor Necrosis Factor Receptor-Associated Peptides and Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D004040:Dietary Carbohydrates; D029941:Fish Proteins; D008055:Lipids; D012333:RNA, Messenger; D047988:Tumor Necrosis Factor Receptor-Associated Peptides and Proteins
keywords: Ctenopharyngodon idella; Immune defense; Inflammation; TRIP (tumor necrosis factor (TNF) receptor-associated factor (TRAF)-Interacting protein; Tissue distribution