Insights Into Determinants That Contribute to Colonization, Virulence and Antibiotic Resistance In Enterococci

<p><em>Enterococcus faecalis</em> and <em>Enterococcus faecium</em> are increasingly common as causative agents of human infections, many of which are very difficult to treat due to multi-drug resistance. The work presented in the first part of this dissertation elucidates a mechanism for the regulation of pilus expression. I showed that ATT is the initiation codon of <em>ebpA</em>, the first gene of an operon that codes for the <em>e</em>ndocarditis and <em>b</em>iofilm-associated <em>p</em>ili (Ebp), a recognized virulence factor in <em>E. faecalis. </em>The presence of this rare start codon downregulates EbpA translation and protein levels, diminishing biofilm and binding abilities of <em>E. faecalis</em>, as compared to an engineered ATG codon. My studies also extended to the ortholog of Ebp in <em>E. faecium</em>, known as the <em>E. faecium</em> <em>p</em>ili (Emp), where the role of each Emp subunit in biofilm formation, adherence and experimental infection was demonstrated. This study highlighted the relevance of the tip subunit, EmpA, in pilus biogenesis and pilus-associated functions. Due to the rising clinical importance of <em>E. faecium</em> and the fact that enterococcal infections are commonly preceded by intestinal colonization, the second part of this dissertation focused on the dynamics of gastrointestinal tract (GIT) colonization of the three known <em>E. faecium</em> clades. This work found that clade B, composed of strains that are part of the normal human microbiota, outcompeted most of the clade A strains, which include strains linked to human infections or associated with animals, when present together in a mouse model of GIT colonization. Last, I demonstrated that the <em>pbp5 </em>gene, part of the <em>E. faecium</em> core genome that shows sequence variation between the clades and encodes a penicillin binding protein important for β-lactam resistance, is differentially expressed between strains, with higher PBP5 protein levels in clade A ampicillin-resistant strains as compared to clade B and subclade A2 ampicillin-susceptible strains. Furthermore, I found evidence that there are extensive differences within the region upstream of <em>pbp5</em> among the clades that correlate with the differential abundance of PBP5 and ampicillin resistance. Together these studies provide further insight into determinants that contribute to colonization, virulence and resistance in enterococci.</p>

Name thesis:degree_name

Doctor of Philosophy (PhD)

Level thesis:degree_level

Dissertation (PhD)

Year dc:date.available

2016

Author dc:creator

• Montealegre, Maria Camila

Contributors dc:contributor

• Barbara E. Murray
• Ambro van Hoof
• Barrett R. Harvey

• Enterococcus faecalis
• Enterococcus faecium
• pili
• gastrointestinal tract colonization
• ampicillin resistance
• Penicillin-Binding Protein 5
• Bacteriology
• Laboratory and Basic Science Research
• Medicine and Health Sciences
• Pathogenic Microbiology

Repository record dc:identifier

https://digitalcommons.library.tmc.edu/utgsbs_dissertations/649

OAI identifier oai:identifier

oai:digitalcommons.library.tmc.edu:utgsbs_dissertations-1691