scholarly journals Glutathionylation ofYersinia pestisLcrV and Its Effects on Plague Pathogenesis

mBio ◽  
2017 ◽  
Vol 8 (3) ◽  
Author(s):  
Anthony Mitchell ◽  
Christina Tam ◽  
Derek Elli ◽  
Thomas Charlton ◽  
Patrick Osei-Owusu ◽  
...  
Keyword(s):  
Yersinia Pestis ◽  
Type Iii Secretion ◽  
Glutathione Synthetase ◽  
Bubonic Plague ◽  
Type Iii ◽  
Codon Substitution ◽  
Mixed Disulfides ◽  
Ribosomal Protein S3 ◽  
Molecular Bases ◽  
Host Tissues

ABSTRACTGlutathionylation, the formation of reversible mixed disulfides between glutathione and protein cysteine residues, is a posttranslational modification previously observed for intracellular proteins of bacteria. Here we show thatYersinia pestisLcrV, a secreted protein capping the type III secretion machine, is glutathionylated at Cys273and that this modification promotes association with host ribosomal protein S3 (RPS3), moderatesY. pestistype III effector transport and killing of macrophages, and enhances bubonic plague pathogenesis in mice and rats. Secreted LcrV was purified and analyzed by mass spectrometry to reveal glutathionylation, a modification that is abolished by the codon substitution Cys273Ala inlcrV. Moreover, thelcrVC273Amutation enhanced the survival of animals in models of bubonic plague. Investigating the molecular mechanism responsible for these virulence attributes, we identified macrophage RPS3 as a ligand of LcrV, an association that is perturbed by the Cys273Ala substitution. Furthermore, macrophages infected by thelcrVC273Avariant displayed accelerated apoptotic death and diminished proinflammatory cytokine release. Deletion ofgshB, which encodes glutathione synthetase ofY. pestis, resulted in undetectable levels of intracellular glutathione, and we used aY. pestisΔgshBmutant to characterize the biochemical pathway of LcrV glutathionylation, establishing that LcrV is modified after its transport to the type III needle via disulfide bond formation with extracellular oxidized glutathione.IMPORTANCEYersinia pestis, the causative agent of plague, has killed large segments of the human population; however, the molecular bases for the extraordinary virulence attributes of this pathogen are not well understood. We show here that LcrV, the cap protein of bacterial type III secretion needles, is modified by host glutathione and that this modification contributes to the high virulence ofY. pestisin mouse and rat models for bubonic plague. These data suggest thatY. pestisexploits glutathione in host tissues to activate a virulence strategy, thereby accelerating plague pathogenesis.

Yersinia pestis outer membrane type III secretion protein YscC: expression, purification, characterization, and induction of specific antiserum

2005 ◽  
Vol 40 (1) ◽  
pp. 152-163 ◽  
Author(s):  
Jeremy L. Goodin ◽  
Ronald W. Raab ◽  
Robert L. McKown ◽  
George L. Coffman ◽  
Bradford S. Powell ◽  
...  
Keyword(s):  
Yersinia Pestis ◽  
Outer Membrane ◽  
Type Iii Secretion ◽  
Specific Antiserum ◽  
Type Iii ◽  
Membrane Type

scholarly journals RfaL Is Required for Yersinia pestis Type III Secretion and Virulence

2013 ◽  
Vol 81 (4) ◽  
pp. 1186-1197 ◽  
Author(s):  
Andrew S. Houppert ◽  
Lesley Bohman ◽  
Peter M. Merritt ◽  
Christopher B. Cole ◽  
Adam J. Caulfield ◽  
...  
Keyword(s):  
Yersinia Pestis ◽  
Type Iii Secretion ◽  
Feedback Inhibition ◽  
Regulatory Gene ◽  
Host Cells ◽  
Proper Function ◽  
Mammalian Host ◽  
Content Type ◽  
Type Iii

ABSTRACTYersinia pestis, the causative agent of plague, uses a type III secretion system (T3SS) to inject cytotoxic Yop proteins directly into the cytosol of mammalian host cells. The T3SS can also be activatedin vitroat 37°C in the absence of calcium. The chromosomal generfaL(waaL) was recently identified as a virulence factor required for proper function of the T3SS. RfaL functions as a ligase that adds the terminalN-acetylglucosamine to the lipooligosaccharide core ofY. pestis. We previously showed that deletion ofrfaLprevents secretion of Yopsin vitro. Here we show that the divalent cations calcium, strontium, and magnesium can partially or fully rescue Yop secretionin vitro, indicating that the secretion phenotype of therfaLmutant may be due to structural changes in the outer membrane and the corresponding feedback inhibition on the T3SS. In support of this, we found that the defect can be overcome by deleting the regulatory genelcrQ. Consistent with a defective T3SS, therfaLmutant is less virulent than the wild type. We show here that the virulence defect of the mutant correlates with a decrease in both T3SS gene expression and ability to inject innate immune cells, combined with an increased sensitivity to cationic antimicrobial peptides.

scholarly journals Global Gene Expression Profiling of Yersinia pestis Replicating inside Macrophages Reveals the Roles of a Putative Stress-Induced Operon in Regulating Type III Secretion and Intracellular Cell Division

2010 ◽  
Vol 78 (9) ◽  
pp. 3700-3715 ◽  
Author(s):  
Hana S. Fukuto ◽  
Anton Svetlanov ◽  
Lance E. Palmer ◽  
A. Wali Karzai ◽  
James B. Bliska
Keyword(s):  
Gene Expression ◽  
Yersinia Pestis ◽  
Type Iii Secretion ◽  
Dna Microarrays ◽  
Cell Envelope ◽  
Global Gene Expression ◽  
Reading Frame ◽  
Type Iii ◽  
Global Gene Expression Profiling ◽  
High Level

ABSTRACT Yersinia pestis, the causative agent of plague, is a facultative intracellular pathogen. Previous studies have indicated that the ability of Y. pestis to survive inside macrophages may be critical during the early stages of plague pathogenesis. To gain insights into the biology of intracellular Y. pestis and its environment following phagocytosis, we determined the genome-wide transcriptional profile of Y. pestis KIM5 replicating inside J774.1 macrophage-like cells using DNA microarrays. At 1.5, 4, and 8 h postinfection, a total of 801, 464, and 416 Y. pestis genes were differentially regulated, respectively, compared to the level of gene expression of control bacteria grown in tissue culture medium. A number of stress-response genes, including those involved in detoxification of reactive oxygen species, as well as several metabolic genes involved in macromolecule synthesis, were significantly induced in intracellular Y. pestis, consistent with the presence of oxidative stress and nutrient starvation inside Yersinia-containing vacuoles. A putative stress-induced operon consisting of y2313, y2315, and y2316 (y2313-y2316), and a previously unidentified open reading frame, orfX, was studied further on the basis of its high level of intracellular expression. Mutant strains harboring either deletion, Δy2313-y2316 or ΔorfX, exhibited diverse phenotypes, including reduced effector secretion by the type III secretion system, increased intracellular replication, and filamentous morphology of the bacteria growing inside macrophages. The results suggest a possible role for these genes in regulating cell envelope characteristics in the intracellular environment.

scholarly journals YopK regulates the Yersinia pestis type III secretion system from within host cells

2011 ◽  
Vol 79 (6) ◽  
pp. 1445-1461 ◽  
Author(s):  
Rebecca Dewoody ◽  
Peter M. Merritt ◽  
Andrew S. Houppert ◽  
Melanie M. Marketon
Keyword(s):  
Yersinia Pestis ◽  
Type Iii Secretion ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Host Cells ◽  
Type Iii

scholarly journals Genome-Wide Analysis of Small RNAs Expressed by Yersinia pestis Identifies a Regulator of the Yop-Ysc Type III Secretion System

2014 ◽  
Vol 196 (9) ◽  
pp. 1659-1670 ◽  
Author(s):  
C. A. Schiano ◽  
J. T. Koo ◽  
M. J. Schipma ◽  
A. J. Caulfield ◽  
N. Jafari ◽  
...  
Keyword(s):  
Yersinia Pestis ◽  
Type Iii Secretion ◽  
Small Rnas ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Genome Wide Analysis ◽  
Type Iii ◽  
Genome Wide

scholarly journals A Type III Secretion System Inhibitor Targets YopD while Revealing Differential Regulation of Secretion in Calcium-Blind Mutants of Yersinia pestis

2013 ◽  
Vol 58 (2) ◽  
pp. 839-850 ◽  
Author(s):  
Danielle L. Jessen ◽  
David S. Bradley ◽  
Matthew L. Nilles
Keyword(s):  
Yersinia Pestis ◽  
Type Iii Secretion ◽  
Yersinia Pseudotuberculosis ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Secretion Systems ◽  
Content Type ◽  
Type Iii ◽  
Regulatory Processes ◽  
Secretion Inhibition

ABSTRACTNumerous Gram-negative pathogens rely upon type III secretion (T3S) systems to cause disease. Several small-molecule inhibitors of the type III secretion systems have been identified; however, few targets of these inhibitors have been elucidated. Here we report that 2,2′-thiobis-(4-methylphenol) (compound D), inhibits type III secretion inYersinia pestis,Yersinia pseudotuberculosis, andPseudomonas aeruginosa. YopD, a protein involved in the formation of the translocon and regulatory processes of the type III secretion system, appears to play a role in the inhibition of secretion by compound D. The use of compound D in T3S regulatory mutants demonstrated a difference in secretion inhibition in the presence and absence of calcium. Interestingly, compound D was effective only under conditions without calcium, indicating that a secretion-active needle structure may be necessary for compound D to inhibit secretion.

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