Genomic and proteomic expression analysis of Leishmania promastigote and amastigote life stages:: The Leishmania genome is constitutively expressed
Genomic and proteomic expression analysis of Leishmania promastigote and amastigote life stages:: The Leishmania genome is constitutively expressed
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DOI：
10.1016/j.molbiopara.2006.11.009
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发表时间：
2007-03-01
影响因子：
1.5
通讯作者：
McMaster, W. Robert
中科院分区：
文献类型：
--
作者：
Leifso, Kirk;Cohen-Freue, Gabriela;McMaster, W. Robert
利什曼原虫是一种原生动物寄生虫，在人类中引起广泛的临床疾病，并在一些国家成为主要的公共卫生风险。利什曼原虫的生命周期由白蛉载体中肠内的细胞外有鞭毛的前鞭毛体阶段和哺乳动物宿主巨噬细胞内的形态学不同的细胞内无鞭毛体阶段组成。本研究利用代表8160个基因的DNA寡核苷酸基因组芯片分析了L.主要的前鞭毛体和病变衍生的无鞭毛体。超过94%的基因在两个生命阶段都有表达。先进的统计分析确定了一个令人惊讶的低程度的差异mRNA表达：1.4%的总基因的无鞭毛体和1.5%的前鞭毛体。这些微阵列结果表明，L。主要基因组基本上在两个生命阶段中组成型表达，并表明利什曼原虫组成型适应于在白蛉载体或巨噬细胞宿主中的存活和复制，其利用针对每种截然不同的环境的适当基因组。利用同位素编码亲和标签（ICAT）技术和质谱技术，对L.婴儿前鞭毛体和无鞭毛体差异表达蛋白。在鉴定的91种不同的蛋白质中，8%在无鞭毛体阶段差异表达，20%在前鞭毛体阶段差异表达，其余72%被认为是组成型表达。通过鉴定以前报道的阶段特异性蛋白质，验证了差异表达，并鉴定了几种无鞭毛体和前鞭毛体新的阶段特异性蛋白质。(c)2006 Elsevier B.V.保留所有权利。
Leishinania are protozoan parasites that cause a wide spectrum of clinical diseases in humans and area major public health risk in several countries. Leishmania life cycle consists of an extracellular flagellated promastigote stage within the midgut of a sandfly vector, and a morphological distinct intracellular amastigote stage within macrophages of a mammalian host. This study reports the use of DNA oligonucleotide genome microarrays representing 8160 genes to analyze the mRNA expression profiles of L. major promastigotes and lesion derived amastigotes. Over 94% of the genes were expressed in both life stages. Advanced statistical analysis identified a surprisingly low degree of differential mRNA expression: 1.4% of the total genes in amastigotes and 1.5% in promastigotes. These microarray results demonstrate that the L. major genome is essentially constitutively expressed in both life stages and suggest that Leishmania is constitutively adapted for survival and replication in either the sandfly vector or macrophage host utilizing an appropriate set of genes for each vastly different environment. Quantitative proteomics, using the isotope coded affinity tag (ICAT) technology and mass spectrometry, was used to identify L. infantum promastigote and axenic amastigote differentially expressed proteins. Of the 91 distinct proteins identified, 8% were differentially expressed in the amastigote stage, 20% were differentially expressed in the promastigote stage, and the remaining 72% were considered constitutively expressed. The differential expression was validated by the identification of previously reported stage specific proteins and identified several amastigote and promastigote novel stage specific proteins. (c) 2006 Elsevier B.V. All rights reserved.