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Dive into the research topics where Stephen J. Libby is active.

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Featured researches published by Stephen J. Libby.


Science | 2008

A Nitric Oxide–Inducible Lactate Dehydrogenase Enables Staphylococcus aureus to Resist Innate Immunity

Anthony R. Richardson; Stephen J. Libby; Ferric C. Fang

Staphylococcus aureus is one of the most successful human pathogens, colonizing 2 billion individuals worldwide and causing invasive infections even in immunocompetent hosts. S. aureus can evade multiple components of host innate immunity, including the antimicrobial radical nitric oxide (NO⚫) produced by activated phagocytes. We show that S. aureus is capable of metabolically adapting to nitrosative stress by expressing an NO⚫-inducible l-lactate dehydrogenase (ldh1, SACOL0222) divergently transcribed from the NO⚫-detoxifying flavohemoglobin (hmp). l-Lactate production allows S. aureus to maintain redox homeostasis during nitrosative stress and is essential for virulence. NO⚫-inducible lactate dehydrogenase activity and NO⚫ resistance distinguish S. aureus from the closely related commensal species S. epidermidis and S. saprophyticus.


Molecular Microbiology | 2005

Co‐regulation of Salmonella enterica genes required for virulence and resistance to antimicrobial peptides by SlyA and PhoP/PhoQ

William Wiley Navarre; Thomas A. Halsey; Don Walthers; Jonathan G. Frye; Michael McClelland; Jennifer L. Potter; Linda J. Kenney; John S. Gunn; Ferric C. Fang; Stephen J. Libby

Analysis of the transcriptome of slyA mutant Salmonella enterica serovar Typhimurium revealed that many SlyA‐dependent genes, including pagC, pagD, ugtL, mig‐14, virK, phoN, pgtE, pipB2, sopD2, pagJ and pagK, are also controlled by the PhoP/PhoQ regulatory system. Many SlyA‐ and PhoP/PhoQ‐co‐regulated genes have functions associated with the bacterial envelope, and some have been directly implicated in virulence and resistance to antimicrobial peptides. Purified His‐tagged SlyA binds to the pagC and mig‐14 promoters in regions homologous to a previously proposed ‘SlyA‐box’. The pagC promoter lacks a consensus PhoP binding site and does not bind PhoP in vitro, suggesting that the effect of PhoP on pagC transcription is indirect. Stimulation of pagC expression by PhoP requires SlyA. Levels of SlyA protein and mRNA are not significantly changed under low‐magnesium PhoP‐inducing conditions in which pagC expression is profoundly elevated, however, indicating that the PhoP/PhoQ system does not activate pagC expression by altering SlyA protein concentration. Models are proposed in which PhoP may control SlyA activity via a soluble ligand or SlyA may function as an anti‐repressor to allow PhoP activation. The absence of almost all SlyA‐activated genes from the Escherichia coli K12 genome suggests that the functional linkage between the SlyA and PhoP/PhoQ regulatory systems arose as Salmonella evolved its distinctive pathogenic lifestyle.


Journal of Clinical Investigation | 1995

DNA repair is more important than catalase for Salmonella virulence in mice.

Nancy A. Buchmeier; Stephen J. Libby; Yisheng Xu; Peter C. Loewen; Jacek Switala; Donald G. Guiney; Ferric C. Fang

Pathogenic microorganisms possess antioxidant defense mechanisms for protection from reactive oxygen metabolites such as hydrogen peroxide (H2O2), which are generated during the respiratory burst of phagocytic cells. These defense mechanisms include enzymes such as catalase, which detoxify reactive oxygen species, and DNA repair systems which repair damage resulting from oxidative stress. To determine the relative importance of these two potentially protective defense mechanisms against oxidative stress encountered by Salmonella during infection of the host, a Salmonella typhimurium double mutant unable to produce either the HPI or HPII catalase was constructed, and compared with an isogenic recA mutant deficient in DNA repair. The recA mutant was hypersusceptible to H2O2 at low cell densities in vitro, while the catalase mutant was more susceptible to high H2O2 concentrations at high cell densities. The catalase mutant was found to be resistant to macrophages and retained full murine virulence, in contrast to the recA mutant which previously was shown to be macrophage-sensitive and attenuated in mice. These observations suggest that Salmonella is subjected to low concentrations of H2O2 while at relatively low cell density during infection, conditions requiring an intact DNA repair system but not functional catalase activity.


Infection and Immunity | 2004

The Ferritin-Like Dps Protein Is Required for Salmonella enterica Serovar Typhimurium Oxidative Stress Resistance and Virulence

Thomas A. Halsey; Andrés Vázquez-Torres; Daniel J. Gravdahl; Ferric C. Fang; Stephen J. Libby

ABSTRACT Resistance to phagocyte-derived reactive oxygen species is essential for Salmonella enterica serovar Typhimurium pathogenesis. Salmonella can enhance its resistance to oxidants through the induction of specific genetic pathways controlled by SoxRS, OxyR, σS, σE, SlyA, and RecA. These regulons can be found in a wide variety of pathogenic and environmental bacteria, suggesting that evolutionarily conserved mechanisms defend against oxidative stress both endogenously generated by aerobic respiration and exogenously produced by host phagocytic cells. Dps, a ferritin-like protein found in many eubacterial and archaebacterial species, appears to protect cells from oxidative stress by sequestering iron and limiting Fenton-catalyzed oxyradical formation. In Escherichia coli and some other bacterial species, Dps has been shown to accumulate during stationary phase in a σS-dependent fashion, bind nonspecifically to DNA, and form a crystalline structure that compacts and protects chromatin from oxidative damage. In the present study, we provide evidence that Dps protects Salmonella from iron-dependent killing by hydrogen peroxide, promotes Salmonella survival in murine macrophages, and enhances Salmonella virulence. Reduced numbers of dps mutant bacteria in the livers and spleens of infected mice are consistent with a role of Dps in protecting Salmonella from oxidative stress encountered during infection.


Molecular Microbiology | 2007

The response regulator SsrB activates expression of diverse Salmonella pathogenicity island 2 promoters and counters silencing by the nucleoid-associated protein H-NS.

Don Walthers; Ronan K. Carroll; William Wiley Navarre; Stephen J. Libby; Ferric C. Fang; Linda J. Kenney

The two‐component system SsrA–SsrB activates expression of a type III secretion system required for replication in macrophages and systemic infection in mice. Here we characterize the SsrB‐dependent regulation of genes within Salmonella pathogenicity island 2 (SPI‐2). Primer extension and DNase I footprinting identified multiple SsrB‐regulated promoters within SPI‐2 located upstream of ssaB, sseA, ssaG and ssaM. We previously demonstrated that ssrA and ssrB transcription is uncoupled. Overexpression of SsrB in the absence of its cognate kinase, SsrA, is sufficient to activate SPI‐2 transcription. Because SsrB requires phosphorylation to relieve inhibitory contacts that occlude its DNA‐binding domain, additional components must phosphorylate SsrB. SPI‐2 promoters examined in single copy were highly SsrB‐dependent, activated during growth in macrophages and induced by acidic pH. The nucleoid structuring protein H‐NS represses horizontally acquired genes; we confirmed that H‐NS is a negative regulator of SPI‐2 gene expression. In the absence of H‐NS, the requirement for SsrB in activating SPI‐2 genes is substantially reduced, suggesting a role for SsrB in countering H‐NS silencing. SsrB activates transcription of multiple operons within SPI‐2 by binding to degenerate DNA targets at diversely organized promoters. SsrB appears to possess dual activities to promote SPI‐2 gene expression: activation of transcription and relief of H‐NS‐mediated repression.


Journal of Bacteriology | 2007

FNR Is a Global Regulator of Virulence and Anaerobic Metabolism in Salmonella enterica Serovar Typhimurium (ATCC 14028s)

Ryan C. Fink; Matthew R. Evans; Steffen Porwollik; Andrés Vázquez-Torres; Jessica Jones-Carson; Bryan Troxell; Stephen J. Libby; Michael McClelland; Hosni M. Hassan

Salmonella enterica serovar Typhimurium must successfully transition the broad fluctuations in oxygen concentrations encountered in the host. In Escherichia coli, FNR is one of the main regulatory proteins involved in O2 sensing. To assess the role of FNR in serovar Typhimurium, we constructed an isogenic fnr mutant in the virulent wild-type strain (ATCC 14028s) and compared their transcriptional profiles and pathogenicities in mice. Here, we report that, under anaerobic conditions, 311 genes (6.80% of the genome) are regulated directly or indirectly by FNR; of these, 87 genes (28%) are poorly characterized. Regulation by FNR in serovar Typhimurium is similar to, but distinct from, that in E. coli. Thus, genes/operons involved in aerobic metabolism, NO. detoxification, flagellar biosynthesis, motility, chemotaxis, and anaerobic carbon utilization are regulated by FNR in a fashion similar to that in E. coli. However, genes/operons existing in E. coli but regulated by FNR only in serovar Typhimurium include those coding for ethanolamine utilization, a universal stress protein, a ferritin-like protein, and a phosphotransacetylase. Interestingly, Salmonella-specific genes/operons regulated by FNR include numerous virulence genes within Salmonella pathogenicity island 1 (SPI-1), newly identified flagellar genes (mcpAC, cheV), and the virulence operon (srfABC). Furthermore, the role of FNR as a positive regulator of motility, flagellar biosynthesis, and pathogenesis was confirmed by showing that the mutant is nonmotile, lacks flagella, is attenuated in mice, and does not survive inside macrophages. The inability of the mutant to survive inside macrophages is likely due to its sensitivity to the reactive oxygen species generated by NADPH phagocyte oxidase.


Proceedings of the National Academy of Sciences of the United States of America | 2010

Humanized nonobese diabetic-scid IL2rγnull mice are susceptible to lethal Salmonella Typhi infection

Stephen J. Libby; Michael A. Brehm; Dale L. Greiner; Leonard D. Shultz; Michael McClelland; Kelly D. Smith; Brad T. Cookson; Joyce E. Karlinsey; Traci L. Kinkel; Steffen Porwollik; Rocío Canals; Lisa A. Cummings; Ferric C. Fang

Salmonella enterica serovar Typhi, the cause of typhoid fever, is host-adapted to humans and unable to cause disease in mice. Here, we show that S. Typhi can replicate in vivo in nonobese diabetic (NOD)-scid IL2rγnull mice engrafted with human hematopoietic stem cells (hu-SRC-SCID mice) to cause a lethal infection with pathological and inflammatory cytokine responses resembling human typhoid. In contrast, S. Typhi does not exhibit net replication or cause illness in nonengrafted or immunocompetent control animals. Screening of transposon pools in hu-SRC-SCID mice revealed both known and previously unknown Salmonella virulence determinants, including Salmonella Pathogenicity Islands 1, 2, 3, 4, and 6. Our observations indicate that the presence of human immune cells allows the in vivo replication of S. Typhi in mice. The hu-SRC-SCID mouse provides an unprecedented opportunity to gain insights into S. Typhi pathogenesis and devise strategies for the prevention of typhoid fever.


Infection and Immunity | 2004

The Salmonella enterica Serovar Typhimurium Divalent Cation Transport Systems MntH and SitABCD Are Essential for Virulence in an Nramp1G169 Murine Typhoid Model

Michelle L. Zaharik; Vivian Li Cullen; Angela M. Fung; Stephen J. Libby; Sonya L. Kujat Choy; Bryan Coburn; David G. Kehres; Michael E. Maguire; Ferric C. Fang; B. Brett Finlay

ABSTRACT Nramp1 is a transporter that pumps divalent cations from the vacuoles of phagocytic cells and is associated with the innate resistance of mice to diverse intracellular pathogens. We demonstrate that sitA and mntH, genes encoding high-affinity metal ion uptake systems in Salmonella enterica serovar Typhimurium, are upregulated when Salmonella is internalized by Nramp1-expressing macrophages and play an essential role in systemic infection of congenic Nramp1-expressing mice.


Journal of Clinical Microbiology | 2005

Liver Abscess Caused by magA+ Klebsiella pneumoniae in North America

Ferric C. Fang; Netanya Sandler; Stephen J. Libby

ABSTRACT Taiwan has witnessed an emerging syndrome of liver abscess caused by Klebsiella pneumoniae carrying the magA gene required for exopolysaccharide web biosynthesis. We report a patient transferred from Alaska to Washington State with a magA+K. pneumoniae liver abscess and describe a simple approach for recognition of these hypervirulent strains.


Infection and Immunity | 2005

Isocitrate Lyase (AceA) Is Required for Salmonella Persistence but Not for Acute Lethal Infection in Mice

Ferric C. Fang; Stephen J. Libby; Margaret Castor; Angela M. Fung

ABSTRACT Isocitrate lyase is required for fatty acid utilization via the glyoxylate shunt. Although isocitrate lyase is essential for Salmonella persistence during chronic infection, it is dispensable for acute lethal infection in mice. Substrate availability in the phagosome appears to evolve over time, with increasing fatty acid dependence during chronic infection.

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Ferric C. Fang

University of Washington

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Joshua Fierer

University of California

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