Transcription Regulation in Pseudomonas aeruginosa

铜绿假单胞菌是一种重要的人类机会致病菌，因其 囊性纤维化患者发病和死亡的主要原因；囊性纤维化肺的慢性定植 由铜绿假单胞菌引起的肺炎通常会导致进行性肺损伤，最终导致呼吸衰竭和死亡。 这项拟议的工作将阐明TeX的功能和作用机制，TeX是一种假定的转录因子 来自铜绿假单胞菌，在慢性病模型中是宿主肺部感染所必需的。 TeX在细菌中高度保守，是真核转录延伸因子的同源物 Spt6.这项研究的基本假设是tex在铜绿假单胞菌中扮演着重要的角色。 致病机制是通过影响毒力基因的表达，这些毒力基因本身对感染沙门氏菌很重要， 或者在宿主体内生存。为了支持这一观点，初步实验表明，TeX与 RNA聚合酶(RNAP)是基因表达的中心酶，而TeX可以影响基因的表达 铜绿假单胞菌基因的一个子集。此外，这些实验表明，特克斯不仅与 与RNAP，但也与假定的多亚单位复合体的成分，称为RNA降解体：A 首次在大肠杆菌中描述的含有核糖核酸酶的复合体，据预测主要参与 M RNA的降解。这些发现提出了转录机制可能是 在操作上与铜绿假单胞菌的RNA降解机制有关。可以想象，这样的联系可以 便于对特定文字稿进行任何必要的处理或降级，并可代表小说 引发基因控制的机制。 拟议的研究将识别其表达受TeX影响的基因，识别共同净化的RNA 并测试特定的模型，以确定Tex如何影响基因表达。具有高分辨率的 以Tex的晶体结构为指导，我们将探索Tex的哪些结构特征对其 功能。此外，我们还将进一步研究那些含有tex和tex的蛋白质复合体的性质。 研究TeX与RNAP和P. 铜绿假单胞菌。 拟议的实验不仅应该阐明TeX在人类基因组中的重要性的分子基础 发病机制，但也可能促进开发新的治疗剂，可用于 对抗肺组织铜绿假单胞菌感染。

Pseudomonas aeruginosa is an important opportunistic pathogen of humans that is notorious for being the principal cause of morbidity and mortality in Cystic Fibrosis (CF) patients; chronic colonization of the CF lung by P. aeruginosa typically leads to progressive lung damage, and eventually respiratory failure and death. The proposed work will elucidate the function and mechanism of action of Tex, a putative transcription factor from P. aeruginosa that is required for infection of the host lung in a chronic disease model. Tex is highly conserved amongst bacteria, and is a homolog of the eukaryotic transcription elongation factor Spt6. The underlying hypothesis for the proposed study is that Tex plays an important role in P. aeruginosa pathogenesis by influencing the expression of virulence genes that are themselves important for infection of, or survival within, the host. In support of this idea, preliminary experiments indicate that Tex associates with RNA polymerase (RNAP), the central enzyme of gene expression, and that Tex can influence the expression of a subset of genes in P. aeruginosa. Furthermore these experiments reveal that Tex associates not only with RNAP but also with components of a putative multi-subunit complex, termed the RNA degradosome: a ribonuclease-containing complex first described in E. coli that is predicted to be primarily involved in the degradation of mRNA. These findings raise the possibility that the transcription machinery may be operationally linked to the RNA degradation machinery in P. aeruginosa. Such a link could conceivably facilitate any requisite processing or degradation of a particular transcript and may represent a novel mechanism for eliciting gene control. The proposed studies will identify genes whose expression is influenced by Tex, identify RNAs that co-purify with Tex, and test specific models for how Tex might influence gene expression. With a high-resolution crystal structure of Tex as a guide, we will explore which structural features of Tex are important for its function. Moreover, we will investigate further the nature of those protein complexes that contain Tex and investigate the interactions of Tex with both RNAP and components of the putative degradosome in P. aeruginosa. The proposed experiments should not only shed light on the molecular basis for the importance of Tex in pathogenesis, but may also facilitate the development of novel therapeutic agents that can be used to combat P. aeruginosa infection in the CF lung.