Yeast functional genomic screens lead to identification of a role for a bacterial effector in innate immunity regulation.
Yeast functional genomic screens lead to identification of a role for a bacterial effector in innate immunity regulation.
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DOI：
10.1371/journal.ppat.0030021
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发表时间：
2007-02
期刊：
影响因子：
6.7
通讯作者：
Lesser CF
中科院分区：
文献类型：
--
作者：
Kramer RW;Slagowski NL;Eze NA;Giddings KS;Morrison MF;Siggers KA;Starnbach MN;Lesser CF
许多细菌病原体操纵宿主细胞过程以促进感染，并最终通过它们直接注入宿主细胞的蛋白质的作用引起疾病。鉴定这些细菌效应蛋白的作用靶点和分子机制对于理解发病机制至关重要。我们已经开发了一种系统的生物学方法，使用酵母酿酒酵母，可以加快细菌效应蛋白靶向的细胞过程的识别。我们系统地筛选了活酵母单倍体缺失菌株集合中对志贺氏菌III型效应子OspF表达超敏的突变体。结果的统计数据挖掘确定了几个细胞过程，包括细胞壁生物发生，当缺失导致酵母对OspF表达过敏时，细胞壁生物发生受损。微阵列实验表明，OspF的表达导致酵母细胞壁完整性途径调控的基因的逆转调节。酵母细胞壁完整性途径是高度保守的促分裂原活化蛋白激酶（MAPK）信号传导途径，通常响应于细胞壁扰动而活化。总之，这些结果使我们假设，并随后证明，OspF抑制酵母和哺乳动物MAPK信号级联。此外，在小鼠模型中，OspF对MAPK信号传导的抑制与宿主对志贺氏菌感染的先天免疫应答的减弱有关。这些研究展示了酵母系统生物学如何促进致病性细菌效应蛋白的功能表征。许多细菌病原体使用专门的分泌系统将效应蛋白直接递送到宿主细胞中。效应蛋白介导宿主细胞过程的破坏或抑制以促进病原体的存活。虽然这些蛋白质是发病机制的关键要素，但相对较少得到充分表征。它们通常与已知功能的蛋白质缺乏显著的同源性，并且它们在生物学和实践方面对体内研究提出了特殊的挑战。例如，它们的功能往往显得多余或具有协同作用，而产生它们的生物可能是危险的或难以培养的，需要特殊的设施。酵母酿酒酵母（Saccharomycescerevisiae）最近已成为一种识别和功能表征效应蛋白的模型系统。这项工作描述了如何全基因组表型筛选和表达志贺氏菌效应OspF的酵母的mRNA分析导致发现OspF抑制酵母和哺乳动物细胞中的有丝分裂原活化蛋白激酶信号传导。丝裂原活化蛋白激酶信号传导的这种抑制与宿主先天免疫应答的减弱有关。这项研究表明，酵母功能基因组研究如何有助于了解致病效应蛋白。
Numerous bacterial pathogens manipulate host cell processes to promote infection and ultimately cause disease through the action of proteins that they directly inject into host cells. Identification of the targets and molecular mechanisms of action used by these bacterial effector proteins is critical to understanding pathogenesis. We have developed a systems biological approach using the yeast Saccharomyces cerevisiae that can expedite the identification of cellular processes targeted by bacterial effector proteins. We systematically screened the viable yeast haploid deletion strain collection for mutants hypersensitive to expression of the Shigella type III effector OspF. Statistical data mining of the results identified several cellular processes, including cell wall biogenesis, which when impaired by a deletion caused yeast to be hypersensitive to OspF expression. Microarray experiments revealed that OspF expression resulted in reversed regulation of genes regulated by the yeast cell wall integrity pathway. The yeast cell wall integrity pathway is a highly conserved mitogen-activated protein kinase (MAPK) signaling pathway, normally activated in response to cell wall perturbations. Together these results led us to hypothesize and subsequently demonstrate that OspF inhibited both yeast and mammalian MAPK signaling cascades. Furthermore, inhibition of MAPK signaling by OspF is associated with attenuation of the host innate immune response to Shigella infection in a mouse model. These studies demonstrate how yeast systems biology can facilitate functional characterization of pathogenic bacterial effector proteins. Many bacterial pathogens use specialized secretion systems to deliver effector proteins directly into host cells. The effector proteins mediate the subversion or inhibition of host cell processes to promote survival of the pathogens. Although these proteins are critical elements of pathogenesis, relatively few are well characterized. They often lack significant homology to proteins of known function, and they present special challenges, biological and practical, to study in vivo. For example, their functions often appear to be redundant or synergistic, and the organisms that produce them can be dangerous or difficult to culture, requiring special facilities. The yeast Saccharomyces cerevisiae has recently emerged as a model system to both identify and functionally characterize effector proteins. This work describes how genome-wide phenotypic screens and mRNA profiling of yeast expressing the Shigella effector OspF led to the discovery that OspF inhibits mitogen-activated protein kinase signaling in both yeast and mammalian cells. This inhibition of mitogen-activated protein kinase signaling is associated with attenuation of the host innate immune response. This study demonstrates how yeast functional genomic studies can contribute to the understanding of pathogenic effector proteins.
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影响因子：
10.5
作者：
KAMADA, Y;JUNG, US;LEVIN, DE
通讯作者：
LEVIN, DE
影响因子：
30.5
作者：
Arbibe, Laurence;Kim, Dong Wook;Sansonetti, Philippe J.
通讯作者：
Sansonetti, Philippe J.
影响因子：
5.8
作者：
Berriz, GF;King, OD;Roth, FP
通讯作者：
Roth, FP
DOI：
10.1073/pnas.120163297
发表时间：
2000-06-06
影响因子：
11.1
作者：
Datsenko, KA;Wanner, BL
通讯作者：
Wanner, BL
影响因子：
4.4
作者：
d'Hauteville, H;Khan, S;Sansonetti, PJ
通讯作者：
Sansonetti, PJ