title: Molecular characterization and expression analysis of olive flounder (Paralichthys olivaceus) phospholipase C gamma 1 and gamma 2.
abstract: Phospholipase C gamma 1 and gamma 2 (PLCG1 and PLCG2) are influential in modulating Ca2+and diacylglycerol, second messengers involved in tyrosine kinase-dependent signaling, including growth factor activation. Here, we used RACE (rapid amplification of cDNA ends) to clone cDNA encoding PLCG1 (PoPLCG1) and PLCG2 (PoPLCG2) in the olive flounder (Paralichthys olivaceus). The respective 1313 and 1249 amino acid sequences share high identity with human PLCG1 and PLCG2, and contain the following domains: pleckstrin homology (PH), EF-hand, catalytic X and Y, Src homology 2 (SH2), Src homology 3 (SH3), and C2. Phylogenic analysis and sequence comparison of PoPLCG1 and PoPLCG2 with other PLC isozymes showed a close relationship between the two PLCGs, supported by structural analysis. In addition, tissue expression analysis showed that PoPLCG1 was expressed predominantly in the brain, eye, and heart, whereas PoPLCG2 was expressed principally in gills, esophagus, spleen, and kidney. Following stimulation with LPS and Poly I:C, PoPLCG expression was compared with the expression of inflammatory cytokines IL-1β, IL-6, and TNF-α via reverse transcription-PCR and real-time quantitative PCR. Our results suggest that PoPLCG isozymes perform a critical immune function in olive flounder, being active in pathogen resistance and the inflammation process.
authors.affiliation: Department of Aquatic Life Medicine, Pukyong National University, Busan 608-737, South Korea. Electronic address: wsj2215@naver.com.
authors.forename: Soo Ji
authors.initials: SJ
authors.lastname: Woo
authors.affiliation: Gyeongbuk Native Fish Business Center, Uiseong 37366, South Korea. Electronic address: cho6835@korea.kr.
authors.forename: Hyae In
authors.initials: HI
authors.lastname: Jo
authors.affiliation: Department of Biotechnology, Pukyong National University, Busan 608-737, South Korea. Electronic address: hyunghl@pknu.ac.kr.
authors.forename: Hyung Ho
authors.initials: HH
authors.lastname: Lee
authors.affiliation: Department of Aquatic Life Medicine, Pukyong National University, Busan 608-737, South Korea. Electronic address: jkchung@pknu.ac.kr.
authors.forename: Joon Ki
authors.initials: JK
authors.lastname: Chung
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005412:Flatfishes / Q000235:genetics* / Q000276:immunology*; D005786:Gene Expression Regulation; D007113:Immunity, Innate; D008070:Lipopolysaccharides / Q000494:pharmacology; D009928:Organ Specificity; D051966:Phospholipase C gamma / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D011070:Poly I-C / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment / Q000662:veterinary
chemical_list: D018076:DNA, Complementary; D029941:Fish Proteins; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D051966:Phospholipase C gamma; D011070:Poly I-C
keywords: Olive flounder; Paralichthys olivaceus; Phospholipase C gamma 1 (PLCG1); Phospholipase C gamma 2 (PLCG2); cDNA cloning; mRNA expression