The effect of mucus and mucin Pseudomonas aeruginosa phenotype and interaction with host cells (360G-Wellcome-099832_Z_12_Z)

Pseudomonas aeruginosa colonises mucus in the lungs of individuals with CF resulting in chronic respiratory infections. We will examine how the bacteria respond to the presence of mucin/mucus and whether or not this response has aneffect on expression of mucin binding proteins and on virulence. We will collect data on the differential expression of Pseudomonas Outer Membrane Proteins (OMPs) in response to both the presence of mucin and mucus and the interaction with host cells. Effect of co-culture of the organism with mucus secreting cells and with purified mucin on OMP expression will be examined using SDS PAGE and affinity blots. The role of these proteins in mediating interaction with mucin and glycans will be assessed using mucin affinity blotting, neoglycoconjugate arrays and a novel microarray platform. Bioinformatics techniques will be used to predict Pseudomonas OMPs that have lectin-like properties. This will aid us in identifying protein properties andmotifs that are responsible for facilitating the binding of proteins to glycans. We also propose to use global transcriptional analysis of selected strains of P. aeruginosa by RNA seq grown in the presence and absence of mucin. The biological relevance of bioinfromatic findings will be assessed in the lab.