Heme-Based Oxygen Sensors of Commensal, Symbiotic, and Pathogenic Bacteria

dc.contributor.advisorGilles-Gonzalez, Marie-Aldaen
dc.creatorTuckerman, Jason Roberten
dc.description.abstractDirect oxygen sensors are proteins that serve as "on-off switches" to cause reversible and adaptive changes in the activities of other proteins or genes, with great specificity in response to fluctuations in oxygen concentration. The heme-based oxygen sensors are a large class of direct oxygen sensors that feature direct binding of oxygen to a sensory heme-containing domain. This heme-binding region couples to a regulatory domain within the same polypeptide. The types of functionalities controlled by these oxygen-specific switches are diverse, and include the regulation of protein activities, gene expression, and second messenger elaboration. A primary focus of this work was the biochemical characterization of a pair of heme-based oxygen sensors involved in the control of the bacterial second messenger cyclic diguanylic acid (c-di-GMP) in Escherichia coli. We discovered that these enzymes, designated DosC and DosP, serve as a diguanylate cyclase and c-di-GMP phosphodiesterase pair that associate with components of the E. coli RNA degradosome in vivo. Importantly, one member of these degradosomes, PNPase, is a direct, high-affinity target of c-di-GMP. These findings imply that specialized oxygen-sensing degradosomes exist in E. coli. In these oxygen-sensing degradosomes cellular oxygen levels regulate PNPase processing of specific RNA transcripts via c-di-GMP. A secondary focus of this work was the characterization of a novel two-component system in M. tuberculosis involved in the non-replicating persistent phase of this bacterium in a typical TB infection. Here, the activities of two heme-containing histidine kinases, DosT and DevS, were discovered to be inhibited specifically by oxygen. As DosT and DevS are the primary regulators of the dormancy survival regulator (DosR/DevR) transcription factor, these results contributed a molecular explanation for the numerous observations linking oxygen and DevR to the dormancy phenotype of M. tuberculosis seen both in vitro and in vivo. [Keywords: oxygen sensor; c-di-GMP; histidine kinase; signal transduction; Escherichia coli; Bordetella pertussis; Mycobacterium tuberculosis]en
dc.subjectCyclic GMPen
dc.subjectSecond Messenger Systemsen
dc.titleHeme-Based Oxygen Sensors of Commensal, Symbiotic, and Pathogenic Bacteriaen
thesis.degree.disciplineBiological Chemistryen
thesis.degree.grantorGraduate School of Biomedical Sciencesen
thesis.degree.nameDoctor of Philosophyen