Transcriptional Control of Chlamydial Development

描述（由申请人提供）：沙眼衣原体是发展中国家可预防性失明的主要原因，也是世界上最常见的性传播感染。全世界大约10%的社区获得性呼吸道感染是由肺炎衣原体引起的。沙眼衣原体和肺炎衣原体与更严重的疾病后遗症有关，这些后遗症被认为是由与持续感染相关的慢性炎症引起的。细胞因子IFN-？在控制衣原体感染中起核心作用，并在细胞培养模型中显示可诱导持续生长状态。在IFN-?存在下持续生长已被证明可以阻断生物体的正常发育周期并阻止生物体向感染性形式的分化。我们计划研究一个非编码rna家族在发育周期中的作用以及与IFN- ？-诱导持续生长。待验证的假设是，ncrna在发育周期中发挥重要作用，并在IFN-?存在下集中参与与持续生长相关的变化。非编码rna将使用基因间和全基因组平铺微阵列进行鉴定，并将通过Northern分析和转录本定位程序进行确认。确认的非编码rna的作用机制将通过在大肠杆菌中的共表达研究来研究。最后，非编码rna的调控作用将在正常和IFN-？利用微阵列分析、定量基因表达分析和基因表达与蛋白表达联合研究，诱导沙眼衣原体和肺炎衣原体持续生长。这项工作将有助于更全面地了解沙眼衣原体和肺炎衣原体的发育周期，并有助于了解衣原体对人类先天免疫反应的机制。在一个快速发展的研究领域中，调控rna代表了非传统抗菌药物的一类新靶点。

DESCRIPTION (provided by applicant): Chlamydia trachomatis is the leading cause of preventable blindness in developing countries and the most commonly reported sexually transmitted infection worldwide. Chlamydia pneumoniae is the cause of approximately 10% of community-acquired respiratory infections worldwide. C. trachomatis and C. pneumoniae have been implicated in more serious disease sequelae that have been proposed to arise due to chronic inflammatory conditions associated with persistent infections. The cytokine IFN-? plays a central role in controlling chlamydial infections and has been shown to induce a persistent growth state in cell culture models. Persistent growth in the presence of IFN-? has been shown to block the normal developmental cycle of the organisms and prevent the differentiation of the organism to the infectious form. We plan to study the role of a family of non-coding RNAs in the developmental cycle and the changes associated with IFN- ?- induced persistent growth. The hypothesis to be tested is that ncRNAs play an important role in the developmental cycle and are centrally involved in the changes associated with persistent growth in the presence of IFN-?. Non-coding RNAs will be identified using intergenic and genome-wide tiling microarrays and will be confirmed by Northern analysis and transcript mapping procedures. The mechanism of action of the confirmed non-coding RNAs will be studied using co-expression studies in Escherichia coli. Finally, the regulatory roles of non-coding RNAs will be tested during normal and IFN-?-induced persistent growth of C. trachomatis and C. pneumoniae using microarray analysis, quantitative gene expression analysis and combined gene expression and protein expression studies. The proposed work will result in a more complete understanding of the developmental cycle of C. trachomatis and C. pneumoniae and will lead to a mechanistic understanding of the chlamydial response to the human innate immune-response. Regulatory RNAs represent a new class of targets for non-traditional antibacterials in a rapidly developing field of study. Narrative: The pathogenic chlamydiae are responsible for respiratory disease, sexually transmitted disease, blinding trachoma and associated chronic diseases. This project uses genome level analyses to determine the role of a new class of RNA regulators in normal growth and the response of the bacteria to the host's immune system. These regulators control important aspects of the bacteria's response to its environment and their role in normal and chronic forms of infection will be studied.