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Development and validation of a high-throughput whole cell assay to investigate Staphylococcus aureus adhesion to host ligands
- Source :
- Journal of Biological Chemistry. 295:16700-16712
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- 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.
- Subjects :
- 0301 basic medicine
030102 biochemistry & molecular biology
biology
Drug discovery
High-throughput screening
Mutant
Cell Biology
medicine.disease_cause
Biochemistry
Virulence factor
Cell biology
Bacterial adhesin
Fibronectin
03 medical and health sciences
030104 developmental biology
Staphylococcus aureus
Sortase
medicine
biology.protein
Molecular Biology
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 295
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi...........dc90ebed079d21e242f0d9db536a003c