Characterizing the role of YjiS in Salmonella virulence

第一个资助期用于开发一个综合工作流程，将新的和现有的数据集结合起来，目的是编制一份关于人类病原体肠道沙门氏菌鼠伤寒血清型模型中sORF的全球概述，特别强调在细菌毒力中具有推定作用的sORF。管道的设计是普遍适用于其他生物和研究问题，强调其有用的更广泛的研究社区。通过对MgrB活性的新见解以及小DUF 1127结构域蛋白YjiS作为第二资助期深入分析的候选物的优先次序，证明了所提出方法的适用性。我们现在计划使用先进的技术，如双重RNA-seq和核糖体分析，以及经典的分子生物学方法，来鉴定YjiS的调节子和相互作用组以及这种小蛋白对沙门氏菌毒力的影响。在项目结束时，我们将对YjiS在巨噬细胞感染期间调节毒力的作用进行深入分析。我们将从不同的角度来研究这一点，例如YjiS可能参与的途径，以及它在宿主对感染的反应中引起的变化。我们将描述YjiS采用的分子作用机制，并全面了解它如何影响毒力的不同方面。与此同时，我们将开始验证我们新预测的STsORF，并进行初步分析，以确定具有感染表型的STsORF。有了这个，我们将更好地了解沙门氏菌如何利用小蛋白来适应和操纵宿主环境。

The first funding period was used to develop an integrative workflow combining new and existing datasets with the aim to compile a global overview of sORFs in the model human pathogen Salmonella enterica serovar Typhimurium, with special emphasis on sORFs with putative roles in bacterial virulence. The pipeline was designed to be universal applicable to other organisms and research question underlining its usefulness for a broader research community. The applicability of the proposed approach was demonstrated by novel insights into MgrB activity as well as the prioritization of the small DUF1127 domain protein YjiS as candidate for in-depth analysis in the second funding period. We now plan to use advanced techniques such as Dual RNA-seq and ribosome profiling, together with classical molecular biology approaches, to identify the regulon and interactome of YjiS and the impact of this small protein on Salmonella virulence. At the end of the project we will have performed an in-depth analysis of the role of YjiS in regulating virulence during macrophages infection. We will investigate this from different points of view, such as the pathways YjiS might be involved in, and the changes it induces in the host response to infection. We will describe the molecular mechanism of action that YjiS employs, and draw a comprehensive picture of how it affects different aspects of virulence. In parallel, we will initiate validation of our newly predicted STsORFs and perform a preliminary analysis to identify the ones with an infection phenotype. With this, we will have obtained a better picture of how small proteins are used by Salmonella to adapt to and manipulate the host environment.