In Vivo-Expressed Genes of<i>Pasteurella multocida</i>

Hunt, Meredith Lesley; Boucher, David; Boyce, John D.; Adler, Ben

doi:10.1128/iai.69.5.3004-3012.2001

Cited by 36 publications

(35 citation statements)

References 52 publications

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“…In Salmonella enterica serovar Typhimurium, a pta mutant is attenuated in the mouse model of peritonitis (24). Both ackA and pta are upregulated in vivo in Pasteurella multocida (22). A probable role for acetyl phosphate metabolism in extraintestinal, gram-negative pathogenesis is in the indirect effect acetyl phosphate has on the histone-like protein H-NS (21).…”

Section: Discussionmentioning

confidence: 99%

Uropathogenic Escherichia coli CFT073 Is Adapted to Acetatogenic Growth but Does Not Require Acetate during Murine Urinary Tract Infection

et al. 2008

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In vivo accumulation of D-serine by Escherichia coli CFT073 leads to elevated expression of PAP fimbriae and hemolysin by an unknown mechanism. Loss of D-serine catabolism by CFT073 leads to a competitive advantage during murine urinary tract infection (UTI), but loss of both D-and L-serine catabolism results in attenuation. Serine is the first amino acid to be consumed in closed tryptone broth cultures and precedes the production of acetyl phosphate, a high-energy molecule involved in intracellular signaling, and the eventual secretion of acetate. We propose that the colonization defect associated with the loss of serine catabolism is due to perturbations of acetate metabolism. CFT073 grows more rapidly on acetogenic substrates than does E. coli K-12 isolate MG1655. As shown by transcription microarray results, D-serine is catabolized into acetate via the phosphotransacetylase (pta) and acetate kinase (ackA) genes while downregulating expression of acetyl coenzyme A synthase (acs). CFT073 acs, which is unable to reclaim secreted acetate, colonized mouse bladders and kidneys in the murine model of UTI indistinguishably from the wild type. Both pta and ackA are involved in the maintenance of intracellular acetyl phosphate. CFT073 pta and ackA mutants were screened to investigate the role of acetyl phosphate in UTI pathogenesis. Both single mutants are at a competitive disadvantage relative to the wild type in the kidneys but normally colonize the bladder. CFT073 ackA pta was attenuated in both the bladder and the kidneys. Thus, we demonstrate that CFT073 is adapted to acetate metabolism as a result of requiring a proper cycling of the acetyl phosphate pathway for colonization of the upper urinary tract.Urinary tract infections (UTIs) place a significant burden on the United States healthcare system, costing upwards of $2.4 billion per year (28). The majority of women will experience a UTI in their lifetime, and every year there are approximately 6.8 million physician visits, 1.2 million emergency room visits, a quarter million hospitalizations, and thousands of deaths due to complications of UTIs, most often sepsis (10,18,28,47). Escherichia coli is the most commonly isolated causative agent of community-acquired UTIs (10).The transition from residence in the gastrointestinal tract, where uropathogenic Escherichia coli (UPEC) transiently resides, to the urinary tract represents a significant change in environment. While the gastrointestinal tract is densely populated with many different species of bacteria, the bladder is normally a sterile environment yet one that presents significant challenges to bacterial growth. In addition to the cleansing flow of urine, numerous innate and acquired immune factors challenge the growth of UPEC in the urinary tract. The host defense involves phagocytic attack, antimicrobial peptides, complement lytic and opsonizing factors, and reactive oxygen and nitrogen species. In addition, the urinary tract as reflected in urine is limited in nutrients common to the intestinal tract, esp...

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Section: Discussionmentioning

confidence: 99%

Uropathogenic Escherichia coli CFT073 Is Adapted to Acetatogenic Growth but Does Not Require Acetate during Murine Urinary Tract Infection

et al. 2008

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“…Induced gene expression during pig infection by Streptococcus suis (254,255) and mouse infection by Lactobacillus reuteri (285) was analyzed by an erythromycin resistance-based screening. And gene expression by Pasteurella multocida infecting mice was explored using a kanamycin resistance reporter gene (108). Recombinase-based selection.…”

Section: Selection Strategies In Ivetmentioning

confidence: 99%

“…A thiol-disulfide interchange protein, encoded by dsbD, was isolated with IVET from Pasteurella multocida (108). DsbD is a membrane-bound enzyme that is responsible for maintaining DsbC in the reduced state by translocating electrons from thioredoxins in the cytoplasm.…”

Section: Genes Involved In Central Intracellular Metabolismmentioning

confidence: 99%

“…IVET isolation suggests a possible role for pta-encoded phosphotransacetylase during rice root colonization by Pseudomonas stutzeri (224). Also, in Pasteurella multocida, an ack-pta operon was shown to be specifically expressed during infection of mice (108). Interestingly, the Vibrio cholerae pta gene was previously isolated using the STM screening for genes required for colonization of the host intestine, implying that a Vibrio cholerae pta mutant strain is significantly reduced in virulence efficiency (36).…”

Section: Regulatory Genesmentioning

confidence: 99%

“…Several IVET and DFI studies demonstrated the importance of lipoproteins and other outer membrane proteins during life in the host environment (Table 2). For instance, IVET demonstrated the upregulation of two genes (pcp and glpQ), encoding membrane-associated lipoproteins, in Pasteurella multocida during infection of mice (108). Although no enzymatic function could be assigned to Pcp, it was demonstrated that Pcp is surface exposed (148).…”

Section: Genes Involved In Cell Envelope Structure and Modificationmentioning

confidence: 99%

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Unraveling the Secret Lives of Bacteria: Use of In Vivo Expression Technology and Differential Fluorescence Induction Promoter Traps as Tools for Exploring Niche-Specific Gene Expression

Rediers

Rainey

Vanderleyden

et al. 2005

Microbiol Mol Biol Rev

141

120

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A major challenge for microbiologists is to elucidate the strategies deployed by microorganisms to adapt to and thrive in highly complex and dynamic environments. In vitro studies, including those monitoring genomewide changes, have proven their value, but they can, at best, mimic only a subset of the ensemble of abiotic and biotic stimuli that microorganisms experience in their natural habitats. The widely used gene-to-phenotype approach involves the identification of altered niche-related phenotypes on the basis of gene inactivation. However, many traits contributing to ecological performance that, upon inactivation, result in only subtle or difficult to score phenotypic changes are likely to be overlooked by this otherwise powerful approach. Based on the premise that many, if not most, of the corresponding genes will be induced or upregulated in the environment under study, ecologically significant genes can alternatively be traced using the promoter trap techniques differential fluorescence induction and in vivo expression technology (IVET). The potential and limitations are discussed for the different IVET selection strategies and system-specific variants thereof. Based on a compendium of genes that have emerged from these promoter-trapping studies, several functional groups have been distinguished, and their physiological relevance is illustrated with follow-up studies of selected genes. In addition to confirming results from largely complementary approaches such as signature-tagged mutagenesis, some unexpected parallels as well as distinguishing features of microbial phenotypic acclimation in diverse environmental niches have surfaced. On the other hand, by the identification of a large proportion of genes with unknown function, these promoter-trapping studies underscore how little we know about the secret lives of bacteria and other microorganisms

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