Genomic Organization and Isoform-Dependent Expression Patterns of Wap65 genes in Various Tissues during Immune Challenges in the Mud Loach Misgurnus mizolepis
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Genomic organization, including the structural characteristics of 5´-flanking regions of two 65-kDa protein (WAP65) isoform genes associated with warm temperature acclimation, were characterized and their transcriptional responses to immune challenges were examined in the intestine, kidney and spleen of the mud loach (Misgurnus mizolepis; Cypriniformes). Both mud loach Wap65 isoform genes displayed a 10-exon structure that is common to most teleostean Wap65 genes. The two mud loach Wap65 isoforms were predicted to possess various stress- and immune-related transcription factor binding sites in their regulatory regions; however, the predicted motif profiles differed between the two isoforms, and the inflammation-related transcription factor binding motifs, such as NF-κB and CREBP sites, were more highlighted in the Wap65-2 isoform than the Wap65-1 isoform. The results of qRT-PCR indicated that experimental immune challenges using Edwardsiella tarda, lipopolysaccharide or polyI:C induced the Wap65-2 isoform more than Wap65-1 isoform, although modulation patterns in response to these challenges were tissue- and stimulant-dependent. This study confirms that functional diversification between the two mud loach Wap65 isoforms (i.e., closer involvement of Wap65-2 in the acute phase of inflammation and innate immunity) occurs at the mRNA level in multiple tissues, and suggests that such differential modulation patterns between the two isoforms are related to the different transcription factor binding profiles in their regulatory regions.
Abstract
Genomic organization, including the structural characteristics of 5´-flanking regions of two 65-kDa protein (WAP65) isoform genes associated with warm temperature acclimation, were characterized and their transcriptional responses to immune challenges were examined in the intestine, kidney and spleen of the mud loach (Misgurnus mizolepis; Cypriniformes). Both mud loach Wap65 isoform genes displayed a 10-exon structure that is common to most teleostean Wap65 genes. The two mud loach Wap65 isoforms were predicted to possess various stress- and immune-related transcription factor binding sites in their regulatory regions; however, the predicted motif profiles differed between the two isoforms, and the inflammation-related transcription factor binding motifs, such as NF-κB and CREBP sites, were more highlighted in the Wap65-2 isoform than the Wap65-1 isoform. The results of qRT-PCR indicated that experimental immune challenges using Edwardsiella tarda, lipopolysaccharide or polyI:C induced the Wap65-2 isoform more than Wap65-1 isoform, although modulation patterns in response to these challenges were tissue- and stimulant-dependent. This study confirms that functional diversification between the two mud loach Wap65 isoforms (i.e., closer involvement of Wap65-2 in the acute phase of inflammation and innate immunity) occurs at the mRNA level in multiple tissues, and suggests that such differential modulation patterns between the two isoforms are related to the different transcription factor binding profiles in their regulatory regions.
참고문헌 (22)
- Aliza D, "Identification of Wap65, a human homologue of hemopexin as a copper-inducible gene in swordtail fish, Xiphophorus helleri", Fish Physiol Biochm, 34, pp.129~138 (2008)
- Cho YS, "Genomic organization and mRNA expression of manganese superoxide dismutase (Mn-SOD) from Hemibarbus mylodon (Teleostei, Cypriniformes)", Fish Shellfish Immunol, 27, pp.571~576 (2009)
- Cho YS, "Modulation of warmtemperature- acclimation-associated 65-kDa protein genes (Wap 65-1 and Wap65-2) in mud loach (Misgurnus mizolepis, Cypriniformes) liver in response to different stimulatory treatments", Fish Shellfish Immunol, 32, pp.662~669 (2012)
- Dooley H, "Emergence of the acute-phase protein hemopexin in jawed vertebrates", Mol Immunol, 48, pp.147~152 (2010)
- Hayden MS, "Signaling to NF-κB", Genes Dev, 18, pp.2195~2224 (2004)
- Hirayama M, "Primary structures and gene organizations of two types of Wap65 from the pufferfish Takifugu rubripes", Fish Physiol Biochem, 29, pp.211~224 (2003)
- Hirayama M, "The occurrence of two types of hemopexin-like protein in medaka and differences in their affinity to heme", J Exp Biol, 207, pp.1387~1398 (2004)
- Kikuchi K, "The 65-kDa cytosolic protein associated with warm temperature acclimation in goldfish, Carassius auratus", J Comp Physiol B, 163, pp.349~354 (1993)
- Kikuchi K, "The warm temperature acclimation-related 65-kDa protein, Wap65, in goldfish and its gene expression", J Biol Chem, 270, pp.17087~17092 (1995)
- Kikuchi K, "The Wap65 gene expression of goldfish (Carassius auratus) in association with warm water temperature as well as bacterial lipopolysaccharide (LPS)", Fish Physiol Biochem, 17, pp.423~432 (1997)
- Kim DS, "Induction of triploidy in mud loach (Misgurnus mizolepis) and its effect on gonad development and growth", Aquaculture, 120, pp.263~270 (1994)
- Kim YO, "Genetic organization of two types of flounder warm-temperature acclimation-associated 65-kDa protein and their gene expression profiles", Biosci Biotechnol Biochem, 77, pp.2065~2072 (2013)
- Lee SY, "Genomic organization and functional diversification of two warm-temperature-acclimation-associated 65-kDa protein genes in rockbream (Oplegnathus fasciatus; Perciformes)", Fish Shellfish Immunol, 37, pp.11~21 (2014)
- Nakaniwa M, "Genomic sequences encoding two types of medaka hemopexin-like protein Wap65, and their gene expression profiles in embryos", J Exp Biol, 208, pp.1915~1925 (2005)
- Nam YK, "Molecular characterization of hepcidin gene from mud loach (Misgurnus mizolepis; Cypriniformes)", Fish Shellfish Immunol, 31, pp.1251~1258 (2011)
- Sarropoulou E, "Comparative genomics in teleost species: knowledge transfer by linking the genomes of model and on-model fish species", Comp Biochem Physiol D Genom Proteom, 6, pp.92~102 (2011)
- Sarropoulou E, "Evolution of a multifunctional gene: The warm temperature acclimation protein Wap65 in the European seabass Dicentrarchus labrax", Mol Phylogenet Evol, 55, pp.640~649 (2010)
- Schmittgen TD, "Analyzing real-time PCR data by the comparative CT method", Nat Protoc, 3, pp.1101~1108 (2008)
- Sha Z, "The warm temperature acclimation protein Wap65 as an immune response gene: Its duplicates are differentially regulated by temperature and bacterial infections", Mol Immunol, 45, pp.1458~1469 (2008)
- Shi YH, "Molecular cloning of liver Wap65 cDNA in ayu (Plecoglossus altivelis) and mRNA expression changes following Listonella anguillarum infection", Mol Biol Rep, 37, pp.1523~1529 (2010)
- Truksa J, "The role of STAT, AP-1, E-box and TIEG motifs in the regulation of hepcidin by IL-6 and BMP-9: Lessons from human HAMP and murine Hamp1 and Hamp2 gene promoters", Blood Cell Mol Dis, 39, pp.255~262 (2007)
- Wang P, "Oct-1 functions as a sensor for metabolic and stress signals", Islets, 2, pp.46~48 (2010)