The function of NGO2115 in the Neisseria gonorrhoeae oxidative stress resistance

描述（由申请方提供）：本提案的目的是表征淋病奈瑟菌氧化应激和先天免疫防御反应的新组分。这项建议与美国国立卫生研究院在该地区的使命有关。淋病是造成这个国家重大健康负担的原因，它解决了淋病奈瑟菌的一个基本方面。淋病生物学，其在有机体的发病机理中具有暗示。N.淋病是研究细菌抗氧化机制的理想候选者，因为它与人类宿主的氧化环境专性相关。初步的实验已经确定了一个过氧化氢诱导的基因，NG 02115，当突变时，赋予增强的抗N。淋病对过氧化氢的作用，表明这种蛋白质在氧化应激反应中的作用。NG 02115转录本编码一个假定的转录调节因子，该因子被假定控制在氧化应激反应中起作用的靶基因的表达。目标1的主要焦点将是通过鉴定NG 02115靶基因来确定NG 02115如何控制对氧化剂的抗性或敏感性。这一目标将使用定量RT-PCR和转录微阵列技术来实现。目标1的第二个目标是研究N.淋病和脑膜炎奈瑟氏球菌NG 02115编码序列可能具有功能性后果。目的2的总体目标是表征NG 02115及其靶基因对N.淋病氧化应激反应将使用体外杀伤试验检测在NG 02115和NG 02115调节靶标中携带突变的菌株对几种类型的活性氧的敏感性。在这些菌株中积累的大分子氧化损伤的量也将通过定量氧化DNA和蛋白质修饰来确定。总之，这些实验将确定NG 02115及其靶基因响应的特定类型的氧化应激。目标2的最终目的是确定NG 02115和NG 0215靶基因在促进对原代人中性粒细胞的耐药性或敏感性方面的作用。这些中性粒细胞易感性实验将确定NG 02115及其靶基因与先天免疫系统的生理相关组分相互作用的贡献。 相关性：淋病是一种性传播感染，是美国公共卫生负担的主要贡献者。这项建议的目的是描述一种新的机制，通过这种机制，病原体。淋病奈瑟菌在氧化应激和先天免疫细胞的杀伤下存活，因此能够促进人类宿主的疾病。

DESCRIPTION (provided by applicant): The objective of this proposal is to characterize a novel component of the Neisseria gonorrhoeae oxidative stress and innate immunity defense responses. This proposal is relevant to the mission of the NIH in that N. gonorrhoeae is responsible for a significant health burden in this country and that it addresses a fundamental aspect of N. gonorrhoeae biology that has implications in the pathogenesis of the organism. N. gonorrhoeae is an ideal candidate for studying bacterial oxidative resistance mechanisms due to its obligate association with the oxidative environment of the human host. Preliminary experiments have identified a hydrogen peroxide-induced gene, NG02115, that when mutated confers enhanced resistance of N. gonorrhoeae to hydrogen peroxide, indicating a role for this protein in the oxidative stress response. The NG02115 transcript encodes a putative transcriptional regulator that is hypothesized to control expression of target genes that function in the oxidative stress response. The major focus of Aim 1 will be to determine how NG02115 controls resistance or sensitivity to oxidants by identifying NG02115 target genes. This goal will be accomplished using quantitative RT-PCR and transcriptional microarray techniques. A second goal of Aim 1 is to investigate species-specific differences between the N. gonorrhoeae and Neisseria meningitidis NG02115 coding sequences that may have functional consequences. The overall objective of Aim 2 is to characterize the contribution of NG02115 and its target genes to the N. gonorrhoeae oxidative stress response. Strains harboring mutations in NG02115 and NG02115 regulatory targets will be tested using in vitro killing assays for sensitivity to several types of reactive oxygen species. The amount of macromolecular oxidative damage that accumulates in these strains will also be determined by quantifying oxidative DNA and protein modifications. Together, these experiments will identify the specific types of oxidative stress that NG02115 and its target genes respond to. The final objective of Aim 2 will be to determine the contribution of NG02115 and NG0215 target genes in facilitating either resistance or sensitivity to primary human neutrophils. These neutrophil susceptibility experiments will establish the contribution of NG02115 and its target genes to interactions with a physiologically relevant component of the innate immune system. Relevance: Gonorrhea is a sexually transmitted infection that is a major contributor to the public health burden in the United States. This proposal aims to characterize a novel mechanism by which the causative agent. Neisseria gonorrhoeae survives oxidative stress and killing by innate immune cells and thus is able to promote disease within the human host.