Role of a cell wall assembly factor in enterococcal antimicrobial resistance

项目总结 几乎所有的革兰氏阳性细菌都会合成一种跨膜的丝氨酸/苏氨酸激酶，它含有3-5个 控制包括抗生素在内的关键过程的细胞外通心区(‘通心粉激酶’) 抗药性、毒素产生、毒性或细胞分裂；在某些细菌中，这种激酶是 生存能力。因此，这些激酶是新疗法的诱人靶点。然而，一个基本的 了解这个家族中的蛋白激酶感知环境的机制 在体内的刺激和处理的信息，以协调适应性生物反应是缺乏的。 这些信息对于开发新的治疗方法至关重要。这项研究 此处提出的建议旨在通过阐明 在这个家族中具有代表性的激酶，即粪肠球菌中的Irek激酶，我们已经证明是 粪肠球菌对细胞壁活性抗生素和 胆汁，如胆酸盐。这项研究旨在阐明对Irek功能的新见解 研究其与其他细胞壁组装因子的物理和功能关系。 粪便。特别是，我们将定义被认为参与其中的蛋白质复合体的组成。 细胞壁组装，评估其在内在抗菌素耐药性中的作用，并检查其相互作用 了解艾瑞克的身体和功能，了解艾瑞克是如何履行职责的。这部作品具有 在细菌和细菌中建立一种新的通过面食蛋白激酶进行跨膜信号传递的模型的可能性 提供关于抗菌素耐药性、细胞壁完整性的协调、毒力和 革兰氏阳性细菌的其他过程。

PROJECT SUMMARY Nearly all Gram-positive bacteria synthesize a transmembrane, Ser/Thr kinase containing 3-5 extracellular PASTA-domains (‘PASTA kinase’) that controls critical processes including antibiotic resistance, toxin production, virulence, or cell division; in some bacteria this kinase is essential for viability. As such, these kinases represent attractive targets for new therapeutics. However, a basic understanding of the mechanisms by which kinases in this family function to perceive environmental stimuli in vivo and process that information to coordinate adaptive biological responses is lacking. Such information is critical to inform development of new therapeutic approaches. The research proposed here seeks to help address this gap by elucidating fundamental aspects of function for a representative kinase in this family, the IreK kinase in Enterococcus faecalis, which we have shown is required for intrinsic resistance of E. faecalis to cell-wall-active antibiotics and to detergents present in bile, such as cholate. This research is designed to elucidate new insights into the function of the IreK kinase by exploring its physical and functional relationships with other cell wall assembly factors in E. faecalis. In particular, we will define the composition of a protein complex thought to be involved in cell wall assembly, assess its role in intrinsic antimicrobial resistance, and examine its interactions both physical and functional with IreK to understand how IreK carries out its duties. This work has the potential to establish a new model for transmembrane signaling by PASTA kinases in bacteria and provide new insights into antimicrobial resistance, coordination of cell wall integrity, virulence and other processes in Gram-positive bacteria.