Staphylococcus aureus ribosomal protein processing, a potential new drug target

描述（由申请人提供）：金黄色葡萄球菌是一种多用途的人类病原体，可导致卫生保健相关感染和越来越多的社区获得性疾病。金黄色葡萄球菌引起多种疾病，从轻微的皮肤和软组织感染到更严重的疾病，包括肺炎、中毒性休克、骨髓炎和心内膜炎。它通常与留置医疗器械感染和手术伤口有关。对多种抗生素具有耐药性的侵袭性菌株的出现，加上广泛的水平基因转移，构成了重大的公共卫生危害。金黄色葡萄球菌快速适应抗生素压力的能力强调了继续开发抗菌治疗新靶点的必要性。我们实验室最近的工作发现，在金黄色葡萄球菌和其他革兰氏阳性细菌中，核糖体大亚基中的一种蛋白质受到特定n端切割的影响。有几条证据与这个分裂事件是必不可少的假设是一致的。生物信息学和生物化学研究已经确定了负责这种切割的蛋白酶。该建议包括遗传和生化方法的结合，以确认裂解和蛋白酶的表征的要求。我们将开发一种裂解实验，并确定蛋白酶和核糖体蛋白中对这种靶向裂解事件所必需的关键氨基酸残基。这些研究将阐明这种新型核糖体蛋白加工事件的作用，并为随后开发一类抑制这种切割反应的新型抗生素奠定基础。

DESCRIPTION (provided by applicant): Staphylococcus aureus is a versatile human pathogen that is responsible for both healthcare-associated infections and, increasingly, community acquired disease. S. aureus causes a wide range of diseases, from minor skin and soft tissue infections to more serious disorders including pneumonia, toxic shock, osteomyelitis and endocarditis. It is commonly found associated with infection of indwelling medical devices and surgical wounds. The emergence of aggressive strains resistant to multiple antibiotics, in combination with widespread horizontal gene transfer, poses a significant public health hazard. The ability of S. aureus to adapt rapidly to antibiotic pressure underscores the need for continued development of new targets for antimicrobial therapy. Recent work in our laboratory has led to the discovery that one of the proteins in the large subunit of the ribosome is subject t a specific N-terminal cleavage in S. aureus and other Gram positive bacteria. Several lines of evidence are consistent with the hypothesis that this cleavage event is essential. Bioinformatic and biochemical studies have led to the identification of the protease responsible for this cleavage. This proposal includes a combination of genetic and biochemical approaches to confirm the requirement for cleavage and the characterization of the protease. We will develop a cleavage assay and identify key amino acid residues in the protease and in the ribosomal protein that are necessary for this targeted cleavage event. These studies will elucidate the role of this novel ribosomal protein processing event and set the stage for subsequent efforts aimed at the development of a new class of antibiotics that will inhibit this cleavage reaction.