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首页> 外文期刊>Traffic >Function and Mechanism of Action of Dictyostelium Nramp1 (Slc11a1) in Bacterial Infection
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Function and Mechanism of Action of Dictyostelium Nramp1 (Slc11a1) in Bacterial Infection

机译：Dictyostelium Nramp1（Slc11a1）在细菌感染中的功能和作用机理

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摘要

Dictyostelium amoebae are professional phagocytes, which ingest bacteria as the principal source of food. We have cloned the Dictyostelium homologue of human natural resistance-associated membrane protein 1 (Nramp1) [solute carrier family 11 member 1 (Slc11a1)], an endo-lysosomal membrane protein that confers on macrophages resistance to infection by a variety of intracellular bacteria and protozoa. The Dictyostelium Nramp1 gene encodes a protein of 53 kDa with 11 putative transmembrane domains. The Nramp1 gene is transcribed during the growth-phase and downregulated to barely detectable levels upon starvation. To gain insights into their intracellular localization, we fused Nramp1 or the vatB subunit of the V-H~+ATPase with green fluorescent protein and expressed in cells. Green fluorescent protein-vatB was inserted in membranes of all acidic compartments and the contractile vacuole network and decorated macropinosomes and phagosomes. Green fluorescent protein-Nramp1 decorated macropinosomes and phagosomes, in addition to intracellular vesicular compartments positive for endosomal SNARE protein Vti1 or vacuo-lin, a marker of the exocytic pathway. Nramp1 disruption generated mutants that were more permissive hosts than wild-type cells for intracellular growth of Legionella pneu-mophila and Micobacterium avium. Nramp1 overexpression protected cells from L pneumophila infection. Evidence is provided that Nramp1 transports metal cations out of the phagolysosome in an ATP-dependent process and that L. pneumophila and M. avium use different mechanisms to neutralize Nramp1 activity.
机译：变形杆菌属（Dictyostelium amoebae）是专业的吞噬细胞，其摄取细菌作为食物的主要来源。我们已经克隆了人类天然抗性相关膜蛋白1（Nramp1）[溶质载体家族11成员1（Slc11a1）]的Dictyostelium同源物，一种溶酶体膜蛋白，赋予巨噬细胞抵抗各种细胞内细菌和细菌感染的能力。原生动物。 Dictyostelium Nramp1基因编码53 kDa的蛋白质，具有11个假定的跨膜结构域。 Nramp1基因在生长期被转录并在饥饿时被下调至几乎不可检测的水平。为了深入了解其细胞内定位，我们将Nramp1或V-H〜+ ATPase的vatB亚基与绿色荧光蛋白融合并在细胞中表达。将绿色荧光蛋白-vatB插入所有酸性隔室和可收缩的液泡网络的膜中，并修饰大体小球体和吞噬体。绿色荧光蛋白-Nramp1装饰的巨球体和吞噬体，以及对内体SNARE蛋白Vti1或胞外途径标记物vacuum-lin呈阳性的细胞内囊泡隔室。 Nramp1破坏产生的突变体比军团菌肺炎军团菌和鸟分枝杆菌的细胞内生长要强于野生型细胞。 Nramp1的过表达保护细胞免受嗜肺杆菌的感染。有证据表明Nramp1在ATP依赖性过程中将金属阳离子从吞噬溶酶体中运出，而嗜肺乳杆菌和鸟分枝杆菌使用不同的机制来中和Nramp1的活性。

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