Honors College Thesis
 

Study on the Evolution and Expression of the Pyoverdine Receptor FpvA inPseudomonas aeruginosa

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  • Pseudomonas aeruginosa is a ubiquitous gram-negative bacterium that is capable of causing infections in people who are immune compromised. The bacterium can live in a variety of environments due to the variety of virulence factors it can produce. It is well adapted to living in iron-stringent conditions because it can acquire iron by secreting an iron-binding siderophore called pyoverdine. Pyoverdines are transported back into the cell through a ferripyoverdine receptor, FpvA. P. aeruginosa also secretes S-type pyocins, which degrade the DNA of sensitive cells. Interestingly, S- type pyocins gain entry through pyoverdine receptors and may play a role in the evolution of these receptors and their cognate biosynthesis pyoverdine. To study the interaction between pyocins and pyoverdines, we developed an in vitro selection scheme. We grew P. aeruginosa ATCC 27853 in the presence of pyocin S3 lysate in iron-stringent conditions. We utilized a spotting and passaging system to enrich for resistant cells. The passaging approach provides a foundation for future fpvA evolution studies. A pyocin S3-resistant mutant was analyzed using Sanger sequencing and found to have a 10-base pair deletion in fpvAIIb.
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