Huang Xin, Li Wen, Jin Min, Ma Fu-Tong, Huang Ying, Shi Yan-Ru, Zhao Ling-Ling, Feng Jin-Ling, Ren Qian, Wang Wen
Jiangsu Key Laboratory for Biodiversity & Biotechnology, Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China.
State Key Laboratory Breeding Base of Marine Genetic Resource, Third Institute of Oceanography, Oceanography, SOA, Xiamen 361005, China.
Fish Shellfish Immunol. 2016 Apr;51:282-290. doi: 10.1016/j.fsi.2016.01.043. Epub 2016 Feb 23.
As a type of pattern-recognition proteins, lectins perform important functions in the innate immunity of crustaceans, including prawns. Although several reports showed that C-type lectin domain family (CLEC) importantly functions in host-pathogen interactions, limited research has focused on CLEC in Macrobrachium rosenbergii. In the present study, a new single CRD containing CLEC (designated as MrLec) was reported in freshwater prawns, M. rosenbergii. The full-length cDNA of MrLec consisted of 1027 bp with an open reading frame of 801 bp, which encoded a peptide of 266 amino acid residues. Genomic sequence for MrLec was also obtained from the M. rosenbergii, which contain 4 exons and 3 introns. MrLec was found to contain a single carbohydrate-recognition domain with an EPN motif. MrLec was ubiquitously distributed in various tissues of a normal prawn, particularly in the hepatopancreas and gills. MrLec expression in the gills was significantly upregulated after a challenge with Vibrio parahaemolyticus and downregulated at 24 h after MrLec RNA interference (MrLec-RNAi). The expression levels of some AMPs, including antilipopolysaccharide factor 1 (Alf1) and lysozyme 2 (Lyso2), also markedly decreased after MrLec-RNAi. Recombinant MrLec can agglutinate (Ca(2+)-dependent) and bind both Gram-negative and Gram-positive bacteria. Results suggested that MrLec participates in the recognition of invading pathogens and functions in the immune response of prawn against pathogen infections.
作为一种模式识别蛋白,凝集素在包括对虾在内的甲壳类动物的先天免疫中发挥着重要作用。尽管有几份报告表明C型凝集素结构域家族(CLEC)在宿主-病原体相互作用中起着重要作用,但针对罗氏沼虾中CLEC的研究却很有限。在本研究中,在淡水虾罗氏沼虾中报道了一种新的含有单个CRD的CLEC(命名为MrLec)。MrLec的全长cDNA由1027个碱基对组成,开放阅读框为801个碱基对,编码一个由266个氨基酸残基组成的肽段。还从罗氏沼虾中获得了MrLec的基因组序列,其包含4个外显子和3个内含子。发现MrLec含有一个带有EPN基序的单一碳水化合物识别结构域。MrLec在正常对虾的各种组织中广泛分布,尤其在肝胰腺和鳃中。在用副溶血性弧菌攻击后,鳃中MrLec的表达显著上调,而在MrLec RNA干扰(MrLec-RNAi)后24小时表达下调。在MrLec-RNAi后,一些抗菌肽(AMPs)的表达水平,包括抗脂多糖因子1(Alf1)和溶菌酶2(Lyso2),也明显下降。重组MrLec可以(依赖Ca(2+))凝集并结合革兰氏阴性菌和革兰氏阳性菌。结果表明,MrLec参与对入侵病原体的识别,并在对虾抵抗病原体感染的免疫反应中发挥作用。