Burge Erin J, Gauthier David T, Ottinger Christopher A, Van Veld Peter A
Department of Environmental and Aquatic Animal Health, Virginia Institute of Marine Science, College of William and Mary, Gloucester Point, Virginia 23062, USA.
Infect Immun. 2004 Mar;72(3):1626-36. doi: 10.1128/IAI.72.3.1626-1636.2004.
In mammals, the natural resistance-associated macrophage protein 1 gene, Nramp1, plays a major role in resistance to mycobacterial infections. Chesapeake Bay striped bass (Morone saxatilis) is currently experiencing an epizootic of mycobacteriosis that threatens the health of this ecologically and economically important species. In the present study, we characterized an Nramp gene in this species and obtained evidence that there is induction following Mycobacterium exposure. The striped bass Nramp gene (MsNramp) and a 554-amino-acid sequence contain all the signal features of the Nramp family, including a topology of 12 transmembrane domains (TM), the transport protein-specific binding-protein-dependent transport system inner membrane component signature, three N-linked glycosylation sites between TM 7 and TM 8, sites of casein kinase and protein kinase C phosphorylation in the amino and carboxy termini, and a tyrosine kinase phosphorylation site between TM 6 and TM 7. Phylogenetic analysis most closely grouped MsNramp with other teleost Nramp genes and revealed high sequence similarity with mammalian Nramp2. MsNramp expression was present in all tissues assayed by reverse transcription-PCR. Within 1 day of injection of Mycobacterium marinum, MsNramp expression was highly induced (17-fold higher) in peritoneal exudate (PE) cells compared to the expression in controls. The levels of MsNramp were three- and sixfold higher on days 3 and 15, respectively. Injection of Mycobacterium shottsii resulted in two-, five-, and threefold increases in gene expression in PE cells over the time course. This report is the first report of induction of an Nramp gene by mycobacteria in a poikilothermic vertebrate.
在哺乳动物中,天然抗性相关巨噬细胞蛋白1基因(Nramp1)在抵抗分枝杆菌感染中起主要作用。切萨皮克湾条纹鲈(Morone saxatilis)目前正经历一场分枝杆菌病的流行,这威胁到了这种在生态和经济上都很重要的物种的健康。在本研究中,我们对该物种中的一个Nramp基因进行了表征,并获得证据表明在暴露于分枝杆菌后会出现诱导现象。条纹鲈Nramp基因(MsNramp)及其554个氨基酸的序列包含Nramp家族的所有信号特征,包括12个跨膜结构域(TM)的拓扑结构、转运蛋白特异性结合蛋白依赖性转运系统内膜成分特征、TM 7和TM 8之间的三个N-连接糖基化位点、氨基和羧基末端的酪蛋白激酶和蛋白激酶C磷酸化位点,以及TM 6和TM 7之间的一个酪氨酸激酶磷酸化位点。系统发育分析显示MsNramp与其他硬骨鱼Nramp基因的亲缘关系最近,并揭示了与哺乳动物Nramp2的高度序列相似性。通过逆转录PCR检测发现,MsNramp在所有检测的组织中均有表达。注射海鱼分枝杆菌1天内,与对照组相比,腹膜渗出液(PE)细胞中MsNramp的表达被高度诱导(高17倍)。在第3天和第15天,MsNramp的水平分别高出3倍和6倍。在整个时间过程中,注射肖茨分枝杆菌导致PE细胞中基因表达分别增加2倍、5倍和3倍。本报告是关于分枝杆菌在变温脊椎动物中诱导Nramp基因的首次报道。