Tn5AraOut mutagenesis for the identification of Yersinia pestis genes involved in resistance towards cationic antimicrobial peptides

Guo, James C. Y.; Nair, Manoj Kumar Mohan; Galván, Estela M.; Liu, Shulin; Schifferli, Dieter M.

doi:10.1016/j.micpath.2011.04.010

Cited by 23 publications

(28 citation statements)

References 77 publications

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“…In that study, an feoC mutant had increased resistance to certain antimicrobial peptides. Similar results were obtained by overexpressing feoAB, leading the authors to conclude that FeoC was repressing expression of the feo operon (38). In our hands, the expression of the feoB::lacZ reporter was unaffected by a mutation in feoC (Fig.…”

Section: Resultssupporting

confidence: 85%

“…Based on these data, and the predicted LysR-like winged-helix N-terminal motif (12), Guo et al proposed that the Y. pestis FeoC is a regulator of feoAB that represses expression in vitro (38). However, our studies using transcriptional reporters failed to show any regulatory effects on transcription from the feo promoter in vitro due to an feoC mutation (Fig.…”

Section: Discussionmentioning

confidence: 73%

“…Recently, Guo et al observed that a transposon insertion into, or deletion of, feoC resulted in increased sensitivity of Y. pestis KIM strains to polymyxin B. Overexpression of feoAB yielded a similar phenotype (38). Based on these data, and the predicted LysR-like winged-helix N-terminal motif (12), Guo et al proposed that the Y. pestis FeoC is a regulator of feoAB that represses expression in vitro (38).…”

Section: Discussionmentioning

confidence: 99%

“…The 9.1-kDa FeoC Yp protein shows the most divergence, being 53% identical and 77% similar to FeoC Ec . Based on bioinformatic analysis and indirect experimental evidence, it has been speculated that FeoC may be a transcriptional repressor of the feoABC locus (7,12,20,23,38,57,66,73,80).…”

mentioning

confidence: 99%

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The Yfe and Feo Transporters Are Involved in Microaerobic Growth and Virulence of Yersinia pestis in Bubonic Plague

et al. 2012

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The Yfe/Sit and Feo transport systems are important for the growth of a variety of bacteria. In Yersinia pestis, single mutations in either yfe or feo result in reduced growth under static (limited aeration), iron-chelated conditions, while a yfe feo double mutant has a more severe growth defect. These growth defects were not observed when bacteria were grown under aerobic conditions or in strains capable of producing the siderophore yersiniabactin (Ybt) and the putative ferrous transporter FetMP. Both fetP and a downstream locus (flp for fet linked phenotype) were required for growth of a yfe feo ybt mutant under static, ironlimiting conditions. An feoB mutation alone had no effect on the virulence of Y. pestis in either bubonic or pneumonic plague models. An feo yfe double mutant was still fully virulent in a pneumonic plague model but had an ϳ90-fold increase in the 50% lethal dose (LD 50 ) relative to the Yfe ؉ Feo ؉ parent strain in a bubonic plague model. Thus, Yfe and Feo, in addition to Ybt, play an important role in the progression of bubonic plague. Finally, we examined the factors affecting the expression of the feo operon in Y. pestis. Under static growth conditions, the Y. pestis feo::lacZ fusion was repressed by iron in a Fur-dependent manner but not in cells grown aerobically. Mutations in feoC, fnr, arcA, oxyR, or rstAB had no significant effect on transcription of the Y. pestis feo promoter. Thus, the factor(s) that prevents repression by Fur under aerobic growth conditions remains to be identified.

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Section: Resultssupporting

confidence: 85%

Section: Discussionmentioning

confidence: 73%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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The Yfe and Feo Transporters Are Involved in Microaerobic Growth and Virulence of Yersinia pestis in Bubonic Plague

et al. 2012

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“…Similar results were reported recently by Guo et al, who showed that the induced expression of the pgbP (also known as pmr) operon in Y. pestis conferred resistance to polymyxin B but not to human cathelicidin LL-37 (23). The reason for this phenotype is unclear, but it appears that the effects of changes in LOS and/or bacterial surface characteristics on Y. pestis sensitivity to antimicrobial peptides are different for cyclic (polymyxin B) versus ␣-helical (cathelicidin) peptides.…”

Section: Discussionsupporting

confidence: 89%

A Transposon Site Hybridization Screen IdentifiesgalUandwecBCas Important for Survival of Yersinia pestis in Murine Macrophages

et al. 2012

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Yersinia pestis is able to survive and replicate within murine macrophages. However, the mechanism by which Y. pestis promotes its intracellular survival is not well understood. To identify genes that are important for Y. pestis survival in macrophages, a library comprised of ϳ31,500 Y. pestis KIM6؉ transposon insertion mutants (input pool) was subjected to negative selection in primary murine macrophages. Genes underrepresented in the output pool of surviving bacteria were identified by transposon site hybridization to DNA oligonucleotide microarrays. The screen identified several genes known to be important for survival of Y. pestis in macrophages, including phoPQ and members of the PhoPQ regulon (e.g., pmrF). In addition, genes predicated to encode a glucose-1-phosphate uridylyltransferase (galU), a UDP-N-acetylglucosamine 2-epimerase (wecB) and a UDP-N-acetyl-D-mannosamine dehydrogenase (wecC) were identified in the screen. Viable-count assays demonstrated that a KIM6؉ galU mutant and a KIM6؉ wecBC mutant were defective for survival in murine macrophages. The galU mutant was studied further because of its strong phenotype. The KIM6؉ galU mutant exhibited increased susceptibility to the antimicrobial peptides polymyxin B and cathelicidin-related antimicrobial peptide (CRAMP). Polyacrylamide gel electrophoresis demonstrated that the lipooligosaccharide (LOS) of the galU mutant migrated faster than the LOS of the parent KIM6؉, suggesting the core was truncated. In addition, the analysis of LOS isolated from the galU mutant by mass spectrometry showed that aminoarabinose modification of lipid A is absent. Therefore, addition of aminoarabinose to lipid A and complete LOS core (galU), as well as enterobacterial common antigen (wecB and wecC), is important for survival of Y. pestis in macrophages.

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Yersinia pestis Transition Metal Divalent Cation Transporters

Perry

Bobrov

Kirillina

et al. 2012

Advances in Yersinia Research

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Tn5AraOut mutagenesis for the identification of Yersinia pestis genes involved in resistance towards cationic antimicrobial peptides

Cited by 23 publications

References 77 publications

The Yfe and Feo Transporters Are Involved in Microaerobic Growth and Virulence of Yersinia pestis in Bubonic Plague

The Yfe and Feo Transporters Are Involved in Microaerobic Growth and Virulence of Yersinia pestis in Bubonic Plague

A Transposon Site Hybridization Screen IdentifiesgalUandwecBCas Important for Survival of Yersinia pestis in Murine Macrophages

Yersinia pestis Transition Metal Divalent Cation Transporters

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