FUNCTIONAL CHARACTERIZATION OF THE LINK BETWEEN CARBOHYDRATE METABOLISM AND THE PATHOGENESIS OF THE INVASIVE M1T1 GROUP A STREPTOCOCCUS

The Group A Streptococcus (GAS), or Streptococcus pyogenes, is a strict human pathogen that colonizes a variety of sites within the host. Infections can vary from minor and easily treatable, to life-threatening, invasive forms of disease. In order to adapt to niches, GAS utilizes environmental cues, such as carbohydrates, to coordinate the expression of virulence factors. Research efforts to date have focused on identifying how either components of the phosphoenolpyruvate-phosphotransferase system (PTS) or global transcriptional networks affect the regulation of virulence factors, but not the synergistic relationship between the two. The present study investigates the role of a putative PTS-fructose operon encoded by fruRBA and its role in virulence in the M1T1 strain 5448. Growth in fructose resulted in induction of fruRBA. RT-PCR showed that fruRBA formed an operon, which was repressed by FruR in the absence of fructose. Growth and carbon utilization profiles revealed that although the entire fruRBA operon was required for growth in fructose, FruA was the main fructose transporter. The ability of both ΔfruR and ΔfruB mutants to survive in whole human blood or neutrophils was impaired. However, the phenotypes were not reproduced in murine whole blood or in a mouse intraperitoneal infection, indicating a human-specific mechanism. While it is known that the PTS can affect activity of the Mga virulence regulator, further characterization of the mechanism by which sugars and its protein domains affect activity have not been studied. Transcriptional studies revealed that the core Mga regulon is activated more in a glucose-rich than a glucose-poor environment. This activation correlates with the differential phosphorylation of Mga at its PTS regulatory domains (PRDs). Using a 5448 mga mutant, transcriptome studies in THY or C media established that the Mga regulon reflects the media used. Interestingly, Mga regulates phage-encoded DNases in a low glucose environment. We also show that Mga activity is dependent on C-terminal amino acid interactions that aid in the formation of homodimers. Overall, the studies presented sought to define how external environmental cues, specifically carbohydrates, control complex regulatory networks used by GAS, contribute to pathogenesis, and aid in adaptation to various nutrient conditions encountered.

Selected Violin Concertos: In Search of a Missing Link

My dissertation presented seven violin concertos in three recital programs. Three of these concertos are acknowledged masterpieces performed in established concert venues throughout the world. They are the concertos of Mozart, Beethoven and Tchaikovsky. The other four are less standard and are composed by Viotti, Kreutzer, Rode and Spohr. These less standard concertos were popular during their time yet they seem not to have stood the test of time. A curriculum devoted exclusively to the standard concertos creates problems for the young violinist. The Mozart violin concertos are often the first standard concertos that the young violin student encounters. They are considered to be the least technically demanding of the standard concertos. The next most advanced standard concertos that the student will usually encounter are Bruch’s G minor concerto, Wieniawski’s D minor concerto and Barber’s concerto. The trouble is that the work on Mozart concertos does not adequately prepare a student for the next most advanced standard concerto. There is a discontinuous leap in the progression of technical difficulty between the Mozart concertos and the next most advanced concertos. Likewise the standard concerto repertoire provides no smooth historical or stylistic progression between the Mozart concertos and the next most advanced concertos. If the young violinist is limited to the standard repertoire then she has no smooth progression either technical, historical or stylistic. I seek to demonstrate that, by adding concertos of Spohr, Viotti, Kreutzer, and Rode to the standard violin curriculum, one could remedy this problem. The first and third recitals were performed in the Gildenhorn Recital Hall and the second recital in the School of Music’s Smith Lecture Hall, both at the University of Maryland. All three recitals can be found in the Digital Repository at the University of Maryland (DRUM).