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Molecular characterization and expressional quantification of lgp2 , a modulatory co‐receptor of RLR‐ signaling pathway in the Indian major carp, Labeo rohita , following pathogenic challenges and PAMPs stimulations
Journal of Fish Biology ( IF 1.7 ) Pub Date : 2020-03-20 , DOI: 10.1111/jfb.14308
Arpita Mohanty 1 , Sushmita Sadangi 1 , Mahismita Paichha 1 , Mrinal Samanta 1
Affiliation  

Lgp2 (laboratory of genetics and physiology 2) is a cytosolic viral sensor of the RLR (retinoic acid-inducible gene 1 like receptor) family member without the caspase recruitment domain (CARD), having both inhibitory and stimulatory role in RLR-signaling pathway. In India, Labeo rohita (rohu) is one of the leading and economically favored freshwater fish species. Several immunological sentry proteins have been reported in this fish species but no information is available on the RLR members. This study was aimed at cloning and characterization of full-length lgp2 cDNA in rohu and its expressional modulations following various PAMPs (pathogen associated molecular patterns) stimulations and bacterial infections. The full- length lgp2-cDNA sequence obtained through RACE (rapid amplifications of cDNA ends)-PCR consisted of 2299 nucleotides with an ORF of 2034 bp encoding 677amino acids. In rohu-Lgp2, four conserved domains viz., a DExDc (DEAD/DEAH box helicase domain), Pfam type III restriction enzyme domain (RES- III), HELICc (Helicase superfamily c-terminal domain) and a Pfam RIG-I_C-RD (RIG-I C-terminal regulatory domain) domain have been detected. Within these domains, several important functional motifs, such as ATP binding site, ATPase motif, RNA unwinding motif and RNA binding sites have also been identified. In healthy rohu, lgp2 gene was abundantly expressed in gill, liver, kidney spleen and blood. In response to both in vitro and in vivo treatment with dsRNA (poly I:C), lgp2 gene expression was significantly (p<0.05) up-regulated in all tested tissues and also in the LRG (Labeo rohita gill) cells. Stimulation of LRG-cells with iE-DAP (γ-D-glutamyl-meso-diaminopimelic acid) and MDP (muramyl dipeptide), lgp2 gene expression was significantly (p<0.05) enhanced. In vivo treatment with LPS and A.hydrophila-derived RNA, resulted in both up and down-regulation of lgp2 gene expression. Upon Gram-positive and Gram-negative bacterial infections, the expression of the lgp2 gene was boosted at different times in almost all the tested tissues. These integrated observations in rohu suggest Lgp2 as an antiviral and antibacterial cytosolic receptor. This article is protected by copyright. All rights reserved.

中文翻译:

在致病性挑战和 PAMP 刺激后,印度主要鲤鱼 Labeo rohita 中 lgp2 的分子表征和表达量化,这是一种调节性共同受体 RLR 信号通路

Lgp2(遗传学和生理学实验室 2)是 RLR(视黄酸诱导基因 1 样受体)家族成员的细胞溶质病毒传感器,没有半胱天冬酶募集结构域 (CARD),在 RLR 信号通路中具有抑制和刺激作用。在印度,Labeo rohita (rohu) 是主要和经济上受青睐的淡水鱼类品种之一。已经在该鱼种中报道了几种免疫哨兵蛋白,但没有关于 RLR 成员的信息。本研究旨在克隆和表征 rohu 中全长 lgp2 cDNA 及其在各种 PAMP(病原体相关分子模式)刺激和细菌感染后的表达调节。通过 RACE(cDNA 末端快速扩增)-PCR 获得的全长 lgp2-cDNA 序列由 2299 个核苷酸组成,ORF 为 2034 bp,编码 677 个氨基酸。在 rohu-Lgp2 中,有四个保守域,即 DExDc(DEAD/DEAH 盒解旋酶域)、Pfam III 型限制酶域(RES-III)、HELICc(解旋酶超家族 c 端域)和 Pfam RIG-I_C- RD(RIG-I C-末端调节域)域已被检测到。在这些域中,还鉴定了几个重要的功能基序,例如 ATP 结合位点、ATP 酶基序、RNA 解旋基序和 RNA 结合位点。在健康罗狐中,lgp2基因在鳃、肝、肾脾和血液中大量表达。响应 dsRNA (poly I:C) 的体外和体内处理,lgp2 基因表达显着(p < 0. 05) 在所有测试组织和 LRG (Labeo rohita gill) 细胞中上调。用 iE-DAP(γ-D-谷氨酰-内消旋二氨基庚二酸)和 MDP(胞壁酰二肽)刺激 LRG 细胞,lgp2 基因表达显着增强(p<0.05)。用 LPS 和 A.hydrophila 衍生的 RNA 进行体内处理,导致 lgp2 基因表达的上调和下调。在革兰氏阳性和革兰氏阴性细菌感染后,lgp2 基因的表达在几乎所有测试组织中的不同时间都得到增强。rohu 中的这些综合观察表明 Lgp2 是一种抗病毒和抗菌细胞溶质受体。本文受版权保护。版权所有。lgp2 基因表达显着增强(p<0.05)。用 LPS 和 A.hydrophila 衍生的 RNA 进行体内处理,导致 lgp2 基因表达的上调和下调。在革兰氏阳性和革兰氏阴性细菌感染后,lgp2 基因的表达在几乎所有测试组织中的不同时间都得到增强。rohu 中的这些综合观察表明 Lgp2 是一种抗病毒和抗菌细胞溶质受体。本文受版权保护。版权所有。lgp2 基因表达显着增强(p<0.05)。用 LPS 和 A.hydrophila 衍生的 RNA 进行体内处理,导致 lgp2 基因表达的上调和下调。在革兰氏阳性和革兰氏阴性细菌感染后,lgp2 基因的表达在几乎所有测试组织中的不同时间都得到增强。rohu 中的这些综合观察表明 Lgp2 是一种抗病毒和抗菌细胞溶质受体。本文受版权保护。版权所有。rohu 中的这些综合观察表明 Lgp2 是一种抗病毒和抗菌细胞溶质受体。本文受版权保护。版权所有。rohu 中的这些综合观察表明 Lgp2 是一种抗病毒和抗菌细胞溶质受体。本文受版权保护。版权所有。
更新日期:2020-03-20
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