Components of SurA Required for Outer Membrane Biogenesis in Uropathogenic Escherichia coli

Background: SurA is a periplasmic peptidyl-prolyl isomerase (PPIase) and chaperone of Escherichia coli and other Gramnegative bacteria. In contrast to other PPIases, SurA appears to have a distinct role in chaperoning newly synthesized porins destined for insertion into the outer membrane. Previous studies have indicated that the chaperone activity of SurA rests in its ‘‘core module’ ’ (the N- plus C-terminal domains), based on in vivo envelope phenotypes and in vitro binding and protection of non-native substrates. Methodology/Principal Findings: In this study, we determined the components of SurA required for chaperone activity using in vivo phenotypes relevant to disease causation by uropathogenic E. coli (UPEC), namely membrane resistance to permeation by antimicrobials and maturation of the type 1 pilus usher FimD. FimD is a SurA-dependent, integral outer membrane protein through which heteropolymeric type 1 pili, which confer bladder epithelial binding and invasion capacity upon uropathogenic E. coli, are assembled and extruded. Consistent with prior results, the in vivo chaperone activity of SurA in UPEC rested primarily in the core module. However, the PPIase domains I and II were not expendable for wild-type resistance to novobiocin in broth culture. Steady-state levels of FimD were substantially restored in the UPEC surA mutant complemented with the SurA N- plus C-terminal domains. The addition of PPIase domain I augmented FimD maturation into the outer membrane, consistent with a model in which domain I enhances stability of and/or substrat

Periplasmic peptidyl-prolyl isomerases SurA and FkpA play an important role in the starvation-stress response (SSR) of Salmonella enterica serovar Typhimurium

Carbon-energy source (C)-starved cells of Salmonella enterica serovar Typhimurium (S. Typhimurium) are remarkably more resistant to stress than actively growing ones. Carbon-starved S. Typhimurium is capable of withstanding extended periods of starvation and assault from a number of different stresses that rapidly kill growing cells. These unique properties of the C-starved cell are the direct result of a series of genetic and physiological adaptations referred to as the starvation-stress response (SSR). Previous work established that the SSR of S. Typhimurium is partially regulated by the extracytoplasmic function sigma factor σE. As part of an effort to identify σE-regulated SSR genes, we investigated surA and fkpA, encoding two different classes of peptidyl-prolyl isomerase that function in folding cell envelope proteins. Both surA and fkpA are members of the heat-shock-inducible σE regulon of Escherichia coli. Although both genes are expressed in C-starved Salmonella cells, evidence indicates that surA and fkpA are not C-starvation-inducible. Furthermore, their expression during C-starvation does not appear to be σE-dependent. Nonetheless, surA and fkpA proved to be important, to differing degrees, for long-term C-starvation survival and for the cross-resistance of C-starved cells to high temperature, acidic pH, and the antimicrobial peptide polymyxin B, but neither were required for cross-resistance to oxidative stress. These results point to fundamental differences between heat-shock-inducible and C-starvation-inducible genes regulated by σE and suggest that genes other than surA and fkpA are involved in the σE-regulated branch of the SSR in Salmonella

Investigation of Yersinia pestis Laboratory Adaptation through a Combined Genomics and Proteomics Approach

The bacterial pathogen Yersinia pestis, the cause of plague in humans and animals, normally has a sylvatic lifestyle, cycling between fleas and mammals. In contrast, laboratory-grown Y. pestis experiences a more constant environment and conditions that it would not normally encounter. The transition from the natural environment to the laboratory results in a vastly different set of selective pressures, and represents what could be considered domestication. Understanding the kinds of adaptations Y. pestis undergoes as it becomes domesticated will contribute to understanding the basic biology of this important pathogen. In this study, we performed a parallel serial passage experiment (PSPE) to explore the mechanisms by which Y. pestis adapts to laboratory conditions, hypothesizing that cells would undergo significant changes in virulence and nutrient acquisition systems. Two wild strains were serially passaged in 12 independent populations each for ~750 generations, after which each population was analyzed using whole-genome sequencing, LC-MS/MS proteomic analysis, and GC/MS metabolomics. We observed considerable parallel evolution in the endpoint populations, detecting multiple independent mutations in ail, pepA, and zwf, suggesting that specific selective pressures are shaping evolutionary responses. Complementary LC-MS/MS proteomic data provide physiological context to the observed mutations, and reveal regulatory changes not necessarily associated with specific mutations, including changes in amino acid metabolism and cell envelope biogenesis. Proteomic data support hypotheses generated by genomic data in addition to suggesting future mechanistic studies, indicating that future whole-genome sequencing studies be designed to leverage proteomics as a critical complement