Staphylococcus aureus adhesion to the host's skin and mucosae enables asymptomatic colonization and the establishment of infection. This process is facilitated by cell wall-anchored adhesins that bind to host ligands. Therapeutics targeting this process could provide significant clinical benefits; however, the development of anti-adhesives requires an in-depth knowledge of adhesion-associated factors and an assay amenable to high-throughput applications. Here, we describe the development of a sensitive and robust whole cell assay to enable the large-scale profiling of S. aureus adhesion to host ligands. To validate the assay, and to gain insight into cellular factors contributing to adhesion, we profiled a sequence-defined S. aureus transposon mutant library, identifying mutants with attenuated adhesion to human-derived fibronectin, keratin, and fibrinogen. Our screening approach was validated by the identification of known adhesion-related proteins, such as the housekeeping sortase responsible for covalently linking adhesins to the cell wall. In addition, we also identified genetic loci that could represent undescribed anti-adhesive targets. To compare and contrast the genetic requirements of adhesion to each host ligand, we generated a S. aureus Genetic Adhesion Network, which identified a core gene set involved in adhesion to all three host ligands, and unique genetic signatures. In summary, this assay will enable high-throughput chemical screens to identify anti-adhesives and our findings provide insight into the target space of such an approach.

中文翻译：

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开发和验证高通量全细胞测定，以研究金黄色葡萄球菌与宿主配体的粘附。


金黄色葡萄球菌粘附在宿主的皮肤和粘膜上，能够无症状定植并建立感染。与宿主配体结合的细胞壁锚定粘附素促进了这一过程。针对这一过程的治疗可以提供显着的临床益处；然而，抗粘连剂的开发需要对粘连相关因素的深入了解和适合高通量应用的检测方法。在这里，我们描述了一种灵敏且稳健的全细胞测定的开发，以能够大规模分析金黄色葡萄球菌与宿主配体的粘附。为了验证该测定，并深入了解有助于粘附的细胞因素，我们分析了序列定义的金黄色葡萄球菌转座子突变体库，鉴定了与人源性纤连蛋白、角蛋白和纤维蛋白原粘附减弱的突变体。我们的筛选方法通过鉴定已知的粘附相关蛋白得到验证，例如负责将粘附素共价连接到细胞壁的管家分选酶。此外，我们还确定了可能代表未描述的抗粘附目标的遗传位点。为了比较和对比与每种宿主配体粘附的遗传要求，我们生成了金黄色葡萄球菌遗传粘附网络，该网络识别了与所有三种宿主配体粘附相关的核心基因集以及独特的遗传特征。总之，该测定将使高通量化学筛选能够识别抗粘合剂，我们的研究结果提供了对这种方法的目标空间的深入了解。