A Genetic Resource for Rapid and Comprehensive Phenotype Screening of Nonessential Staphylococcus aureus Genes

Fey, Paul D.; Endres, Jennifer L.; Yajjala, Vijaya Kumar; Widhelm, Todd J.; Boissy, Robert; Bose, Jeffrey L.; Bayles, Kenneth W.

doi:10.1128/mbio.00537-12

Cited by 788 publications

(882 citation statements)

References 45 publications

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“…The link between c-di-AMP and the ion transport may explain why c-di-AMP, in contrast to other related signaling nucleotides, is essential for growth in bacterial species. Individually ktrA, cpaA, pstA, and kdpD are not essential (28)(29)(30); however, it is plausible that combined mutations may be lethal. Alternatively the existence of an as yet unidentified essential c-di-AMP receptor is also entirely possible.…”

Section: Discussionmentioning

confidence: 99%

Systematic identification of conserved bacterial c-di-AMP receptor proteins

Corrigan

Campeotto

Jeganathan

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

264

405

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Nucleotide signaling molecules are important messengers in key pathways that allow cellular responses to changing environments. Canonical secondary signaling molecules act through specific receptor proteins by direct binding to alter their activity. Cyclic diadenosine monophosphate (c-di-AMP) is an essential signaling molecule in bacteria that has only recently been discovered. Here we report on the identification of four Staphylococcus aureus c-di-AMP receptor proteins that are also widely distributed among other bacteria. Using an affinity pull-down assay we identified the potassium transporter-gating component KtrA as a c-di-AMP receptor protein, and it was further shown that this protein, together with c-di-AMP, enables S. aureus to grow in low potassium conditions. We defined the c-di-AMP binding activity within KtrA to the RCK_C ( r egulator of c onductance of K + ) domain. This domain is also found in a second S. aureus protein, a predicted cation/proton antiporter, CpaA, which as we show here also directly binds c-di-AMP. Because RCK_C domains are found in proteinaceous channels, transporters, and antiporters from all kingdoms of life, these findings have broad implications for the regulation of different pathways through nucleotide-dependent signaling. Using a genome-wide nucleotide protein interaction screen we further identified the histidine kinase protein KdpD that in many bacteria is also involved in the regulation of potassium transport and a P II-like s ignal t ransduction protein, which we renamed PstA, as c-di-AMP binding proteins. With the identification of these widely distributed c-di-AMP receptor proteins we link the c-di-AMP signaling network to a central metabolic process in bacteria.

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Section: Discussionmentioning

confidence: 99%

Systematic identification of conserved bacterial c-di-AMP receptor proteins

Corrigan

Campeotto

Jeganathan

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

264

405

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“…27 Each of these mutants contains a disrupted nonessential gene. These four S. aureus mutants are a Bactericidal assays were done using 25% TSB.…”

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confidence: 99%

Anti-Staphylococcal Biofilm Effects of Human Cathelicidin Peptides

Mishra

Golla

Lau

et al. 2015

ACS Med. Chem. Lett.

101

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Staphylococcus aureus can live together in the form of biofilms to avoid elimination by the host. Thus, a useful strategy to counteract bacterial biofilms is to re-engineer human antimicrobial peptide LL-37 so that it can be used as a remedy for preventing and removing biofilms. This study reports antibiofilm effects of four human cathelicidin LL-37 peptides against community-associated and hospital isolated methicillin-resistant Staphylococcus aureus (MRSA) strains. Although the intact molecule LL-37 inhibited biofilm formation at low concentrations, it did not inhibit bacterial attachment nor disrupt preformed biofilms. However, two 17-residue peptides, GF-17 and 17BIPHE2, inhibited bacterial attachment, biofilm growth, and disrupted established biofilms. An inactive peptide RI-10 was used as a negative control. Our results obtained using the S. aureus mutants in a static biofilm model are consistent with the literature obtained in a flow cell biofilm model. Because 17BIPHE2 is the most effective biofilm disruptor with desired stability to proteases, it is a promising lead for developing new anti-MRSA biofilm agents.

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“…This pattern of insertions is consistent with bias against WTA-associated insertions in the Nebraska Transposon Mutant Library, which consists of 1952 strains, each containing a single mutation within a nonessential gene of S. aureus isolate USA300. This collection also lacks mutations within tagO, tagG and tagH, suggesting these may be essential genes (Fey et al, 2013). Thus the mutations within the hyper-biofilm mutants likely do not knock out WTA synthesis, but may merely reduce it and concomitantly induce sensitivity to bile, leading to an increased level of biofilm production on exposure to lower concentrations of bile.…”

Section: Discussionmentioning

confidence: 99%