Mutations in Three Distinct Loci Cause Resistance to Peptide Deformylase Inhibitors in<i>Bacillus subtilis</i>

Duroc, Yann; Giglione, Carmela; Meinnel, Thierry

doi:10.1128/aac.01340-08

Cited by 16 publications

(18 citation statements)

References 30 publications

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“…In the simplest instances, resistance to an antibacterial agent can arise from mutations that affect its target or its access to or removal from the cell. In the case of PDF inhibitors, mutations that affect formylation of the initiator Met-tRNA, of which those in FMT are the most common, have also been associated with resistance in certain pathogens (12)(13)(14)(15)(16)(17)(18).…”

Section: Discussionmentioning

confidence: 99%

“…In bacteria where protein synthesis can be initiated with unformylated Met-tRNAi (therefore bypassing the need for a deformylation step), the main mechanism of resistance to PDF inhibitors involves loss-of-function mutations in genes involved in the formylation of the initiator tRNA; mutations occur mostly in the gene encoding formyl-methionyl transferase (FMT) (12)(13)(14)(15)(16)(17)(18)(19) but mutations in FolD and GlyA, two enzymes involved in the synthesis of 10-formyl-tetrahydrofolate, have also been described (14,16). Such mutants are highly resistant to PDF inhibitors, but they display compromised in vitro (12,13,(15)(16)(17)(18) and in vivo (12,16,20) growth.…”

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confidence: 99%

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Frequency of Spontaneous Resistance to Peptide Deformylase Inhibitor GSK1322322 in Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus pneumoniae

Min

Ingraham

Huang

et al. 2015

Antimicrob Agents Chemother

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The continuous emergence of multidrug-resistant pathogenic bacteria is compromising the successful treatment of serious microbial infections. GSK1322322, a novel peptide deformylase (PDF) inhibitor, shows good in vitro antibacterial activity and has demonstrated safety and efficacy in human proof-of-concept clinical studies. In vitro studies were performed to determine the frequency of resistance (FoR) to this antimicrobial agent in major pathogens that cause respiratory tract and skin infections. Resistance to GSK1322322 occurred at high frequency through loss-of-function mutations in the formyl-methionyl transferase (FMT) protein in Staphylococcus aureus (4/4 strains) and Streptococcus pyogenes (4/4 strains) and via missense mutations in Streptococcus pneumoniae (6/21 strains), but the mutations were associated with severe in vitro and/or in vivo fitness costs. The overall FoR to GSK1322322 was very low in Haemophilus influenzae, with only one PDF mutant being identified in one of four strains. No target-based mutants were identified from S. pyogenes, and only one or no PDF mutants were isolated in three of the four S. aureus strains studied. In S. pneumoniae, PDF mutants were isolated from only six of 21 strains tested; an additional 10 strains did not yield colonies on GSK1322322-containing plates. Most of the PDF mutants characterized from those three organisms (35/37 mutants) carried mutations in residues at or in close proximity to one of three highly conserved motifs that are part of the active site of the PDF protein, with 30 of the 35 mutations occurring at position V71 (using the S. pneumoniae numbering system).

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Frequency of Spontaneous Resistance to Peptide Deformylase Inhibitor GSK1322322 in Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus pneumoniae

Min

Ingraham

Huang

et al. 2015

Antimicrob Agents Chemother

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“…While mutations in FolD and GlyA, two enzymes involved in the synthesis of 10-formyl-tetrahydrofolate, have been described (13,14), loss-of-function mutations in the gene encoding formylmethionyl transferase (FMT), the enzyme that catalyzes the formylation of the initiator methionyl-tRNA, are the most common cause of resistance to PDF inhibitors in bacteria where FMT is not essential for viability, such as Staphylococcus aureus (3,15), Bacillus subtilis (13), Pseudomonas aeruginosa (16), Salmonella enterica (14), and E. coli (15,17,18). In those organisms, protein synthesis can still initiate with unformylated methionyl-tRNA, bypassing the need for PDF function.…”

mentioning

confidence: 99%

Staphylococcus aureus Formyl-Methionyl Transferase Mutants Demonstrate Reduced Virulence Factor Production and Pathogenicity

Lewandowski

Huang

Fan

et al. 2013

Antimicrob Agents Chemother

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Inhibitors of peptide deformylase (PDF) represent a new class of antibacterial agents with a novel mechanism of action. Mutations that inactivate formyl methionyl transferase (FMT), the enzyme that formylates initiator methionyl-tRNA, lead to an alternative initiation of protein synthesis that does not require deformylation and are the predominant cause of resistance to PDF inhibitors in Staphylococcus aureus. Here, we report that loss-of-function mutations in FMT impart pleiotropic effects that include a reduced growth rate, a nonhemolytic phenotype, and a drastic reduction in production of multiple extracellular proteins, including key virulence factors, such as ␣-hemolysin and Panton-Valentine leukocidin (PVL), that have been associated with S. aureus pathogenicity. Consequently, S. aureus FMT mutants are greatly attenuated in neutropenic and nonneutropenic murine pyelonephritis infection models and show very high survival rates compared with wild-type S. aureus. These newly discovered effects on extracellular virulence factor production demonstrate that FMT-null mutants have a more severe fitness cost than previously anticipated, leading to a substantial loss of pathogenicity and a restricted ability to produce an invasive infection. In bacteria, protein synthesis initiates with formyl-methionyltRNAi, and therefore, all newly synthesized polypeptides contain an N-formyl-methionine terminal end that, in most cases, is not retained in mature proteins. As the polypeptides emerge from the ribosome, the N-formyl group is hydrolyzed by peptide deformylase (PDF), and subsequently, methionine aminopeptidase removes the N-terminal methionine. Deformylation plays a crucial role in protein maturation and has been shown to be essential for bacterial growth (1-4), as methionine aminopeptidase cannot hydrolyze N-blocked peptides (5). The ubiquitous nature of PDF, together with the fact that this function is not required for cytoplasmic protein synthesis in eukaryotes, has made this an attractive target for the development of new antibacterial agents. The successful purification and characterization of the native form of Escherichia coli PDF (6), as well as the discovery that the natural product actinonin is an inhibitor of PDF with weak antibacterial activity (7-9), triggered the search for additional PDF inhibitors. Since then, a large number of structurally diverse PDF inhibitors have been identified, including several compounds with demonstrated in vivo efficacy and good safety profiles (10). Three PDF inhibitors have progressed to clinical trials (11, 12), and one of them, GSK1322322, is in phase II clinical development for the treatment of respiratory tract and skin infections.While mutations in FolD and GlyA, two enzymes involved in the synthesis of 10-formyl-tetrahydrofolate, have been described (13, 14), loss-of-function mutations in the gene encoding formylmethionyl transferase (FMT), the enzyme that catalyzes the formylation of the initiator methionyl-tRNA, are the most common cause of resistance to PDF inhibito...

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“…Single mutations in a gene can change the active amino acid residue, which may play a key role in binding with the drug. Therefore, a drug that has not been strongly bound with amino acids can lead to the appearance of drug resistance [53,54]. To address this issue, we used the 3-D model of PDF for the screening of potent drugs that can be further used for the control of A. hydrophila infection in different animals.…”

Section: The 3-d Modeling Of Peptide Deformylasementioning

confidence: 99%

A Potential Screening of Peptide Deformylase Inhibitors Towards the Control of Aeromonas hydrophila

Singh¹,

Mani²

2017

Curr Synthetic Sys Biol

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Currently, the prevalence of multidrug resistance Aeromonas hydrophila is one of the major issues and challenges for aquatic and terrestrial organisms. Therefore, an urgent need arises to control it using a potent and specific drug. Here, we identified a peptide deformylase (PDF) in A. hydrophila, which is a ubiquitous enzyme and one of the most attractive drug targets. We used the PDF protein sequences for generating a 3-D model using homology modeling. The 3-D model was validated and it was found 91% of the present amino acids in allowed regions of the Ramachandran plot. We used the 3-D model of PDF for the screening of drugs through molecular docking and found BB-3497, actinonin, and BBS-02 were more potent than other studied drugs based on binding energy. We have also generated a phylogenetic tree of PDF from A. hydrophila with other homologous bacteria, suggesting that similar drugs could also be applied to the control of those bacteria. These findings provide a new insight for the better understanding of PDF, which is a novel target for the development of more potent inhibitors towards the better control of multidrug resistant A. hydrophila.

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Mutations in Three Distinct Loci Cause Resistance to Peptide Deformylase Inhibitors inBacillus subtilis

Cited by 16 publications

References 30 publications

Frequency of Spontaneous Resistance to Peptide Deformylase Inhibitor GSK1322322 in Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus pneumoniae

Frequency of Spontaneous Resistance to Peptide Deformylase Inhibitor GSK1322322 in Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus pneumoniae

Staphylococcus aureus Formyl-Methionyl Transferase Mutants Demonstrate Reduced Virulence Factor Production and Pathogenicity

A Potential Screening of Peptide Deformylase Inhibitors Towards the Control of Aeromonas hydrophila

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