Aminoacyl-tRNAs, the bacterial cell envelope, and antibiotics

RajBhandary, Uttam L.; Söll, Dieter

doi:10.1073/pnas.0801193105

Cited by 26 publications

(20 citation statements)

References 27 publications

(23 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The lysyl acceptor and donor substrates turned out to be PG and aminoacyl‐tRNAs (Lennarz et al ., 1966; Nesbitt and Lennarz, 1968), respectively, and these early findings were recently confirmed with recombinant MprF proteins (Staubitz et al ., 2004; Roy and Ibba, 2008, 2009; Klein et al ., 2009). Thus, Lys‐PG biosynthesis belongs to the few examples of non‐ribosomal pathways that employ activated amino acids from aminoacyl tRNAs (RajBhandary and Soll, 2008). MprF represents a new class of transesterase with no detectable relatedness to other transesterases.…”

Section: Aminoacyl Pg Biosynthesis By Mprf Proteinsmentioning

confidence: 99%

Broad‐spectrum antimicrobial peptide resistance by MprF‐mediated aminoacylation and flipping of phospholipids

Ernst

Peschel²

2011

Molecular Microbiology

199

184

View full text Add to dashboard Cite

SummaryBacteria are frequently exposed to cationic antimicrobial peptides (CAMPs) from eukaryotic hosts (host defence peptides) or from prokaryotic competitors (bacteriocins). However, many bacteria, among them most of the major human pathogens, achieve CAMP resistance by MprF, a unique enzyme that modifies anionic phospholipids with L-lysine or L-alanine thereby introducing positive charges into the membrane surface and reducing the affinity for CAMPs. The lysyl or alanyl groups are derived from aminoacyl tRNAs and are usually transferred to phosphatidylglycerol (PG). Recent studies with MprF from Staphylococcus aureus demonstrated that production of Lys-PG only leads to CAMP resistance when an additional flippase domain of MprF is present that translocates Lys-PG and exposes it at the outer leaflet of the membrane. Thus, MprF exerts two specific functions that have hardly been found in other bacterial proteins. MprF proteins are crucial virulence factors of many human pathogens, which recommends them as targets for new anti-virulence drugs. Intriguingly, specific point mutations in mprF cause resistance to the CAMP-like antibiotic daptomycin in a yet unclear way that may involve altered Lys-PG synthesis and/or Lys-PG flipping capacities. Thus, a thorough characterization of MprF domains and functions will help to unravel how bacteria maintain and protect their cytoplasmic membranes.

show abstract

Section: Aminoacyl Pg Biosynthesis By Mprf Proteinsmentioning

confidence: 99%

Broad‐spectrum antimicrobial peptide resistance by MprF‐mediated aminoacylation and flipping of phospholipids

Ernst

Peschel²

2011

Molecular Microbiology

199

184

View full text Add to dashboard Cite

show abstract

“…In contrast, k off was found to remain essentially constant up to an overall lysyl-DOPG content of ϳ30 mol% (Fig. 2C), which encompasses the lysinylation profile of most clinical S. aureus isolates (6,12,13). Only at a lysyl-DOPG concentration of Ͼ30 mol% did k off become notably faster, leading to a significantly reduced net association and a correspondingly higher K D (Fig.…”

mentioning

confidence: 94%

“…For instance, reduced levels of lysyl-PG were found to increase the susceptibility of S. aureus to many antibiotics, including some CAPs (10,18,19), and mutant S. aureus strains that do not produce lysyl-PG exhibit reduced virulence in a variety of in vivo models (18). Of note, the observed correlation between PG lysinylation and reduced susceptibility to cationic antibiotics (e.g., daptomycin) and CAPs has been attributed to a relative decrease in the negative surface charge of the target bacterial cytoplasmic membrane as a consequence of PG modification (8,10,12,13,18). However, no detailed analyses of the mechanism(s) by which lysyl-PG protects the organism from cationic agent-induced killing have been carried out in either whole bacterial cells or model membranes.…”

mentioning

confidence: 99%

“…Esterification of the phosphatidylglycerol (PG) headgroup occurs with either lysine (lysyl-PG), alanine (alanyl-PG), or, less commonly, ornithine (ornithyl-PG) or arginine (arginyl-PG). Such modifications may serve to modulate the cytoplasmic membrane charge, as the adduct converts the anionic PG into a net cationic or zwitterionic lipid, depending on the amino acid involved (13). In Staphylococcus aureus, this reaction is mediated by the multiple peptide resistance factor F (MprF), an integral membrane protein that catalyzes the transfer of the aminoacyl group from Lys-tRNA Lys to the sn-3 position of the PG headgroup (14).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Lysyl-Phosphatidylglycerol Attenuates Membrane Perturbation Rather than Surface Association of the Cationic Antimicrobial Peptide 6W-RP-1 in a Model Membrane System: Implications for Daptomycin Resistance

Kilelee

Pokorny

Yeaman

et al. 2010

Antimicrob Agents Chemother

View full text Add to dashboard Cite

The presence of the cationic phospholipid lysyl-phosphatidylglycerol (lysyl-PG) in staphylococcal cytoplasmic membranes has been linked to increased resistance to cationic compounds, including antibiotics such as daptomycin as well as host defense antimicrobial peptides. We investigated the effects of lysyl-PG on binding of 6W-RP-1, a synthetic antimicrobial peptide, to lipid vesicles and on peptide-induced membrane permeabilization. Unexpectedly, physiological lysyl-PG concentrations only minimally reduced membrane binding of 6W-RP-1. In contrast, 6W-RP-1-induced dye leakage was severely inhibited by lysyl-PG, suggesting that lysyl-PG primarily impacts membrane defect formation.

show abstract

“…In vivo activities of Streptomyces SerRS1 and SerRS2. serS1, serS2, E. coli serS, and the serS2(H270G) mutant were cloned into pUC19 (pUC) vector under the control of the glnS Ј promoter (25). (A) The plasmids were transformed into an E. coli temperature-sensitive (ts) mutant strain K28 that cannot produce chromosomal SerRS at 40°C.…”

Section: Sav4244 Is From Streptomyces Avermitilis (Genbank Accession mentioning

confidence: 99%

Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974

Zeng

Roy

Patil

et al. 2009

Antimicrob Agents Chemother

View full text Add to dashboard Cite

The Trojan horse antibiotic albomycin, produced by Streptomyces sp. strain ATCC 700974, contains a thioribosyl nucleoside moiety linked to a hydroxamate siderophore through a serine residue. The seryl nucleoside structure (SB-217452) is a potent inhibitor of seryl-tRNA synthetase (SerRS) in the pathogenic bacterium Staphylococcus aureus, with a 50% inhibitory concentration (IC 50 ) of ϳ8 nM. In the albomycinproducing Streptomyces sp., a bacterial SerRS homolog (Alb10) was found to be encoded in a biosynthetic gene cluster in addition to another serRS gene (serS1) at a different genetic locus. Alb10, named SerRS2 herein, is significantly divergent from SerRS1, which shows high homology to the housekeeping SerRS found in other Streptomyces species. We genetically and biochemically characterized the two genes and the proteins encoded. Both genes were able to complement a temperature-sensitive serS mutant of Escherichia coli and allowed growth at a nonpermissive temperature. serS2 was shown to confer albomycin resistance, with specific amino acid residues in the motif 2 signature sequences of SerRS2 playing key roles. SerRS1 and SerRS2 are comparably efficient in vitro, but the K m of serine for SerRS2 measured during tRNA aminoacylation is more than 20-fold higher than that for SerRS1. SB-217452 was also enzymatically generated and purified by two-step chromatography. Its IC 50 against SerRS1 was estimated to be 10-fold lower than that against SerRS2. In contrast, both SerRSs displayed comparable inhibition kinetics for serine hydroxamate, indicating that SerRS2 was specifically resistant to SB-217452. These data suggest that mining Streptomyces genomes for duplicated aminoacyltRNA synthetase genes could provide a novel approach for the identification of natural products targeting aminoacyl-tRNA synthetases.

show abstract

Aminoacyl-tRNAs, the bacterial cell envelope, and antibiotics

Cited by 26 publications

References 27 publications

Broad‐spectrum antimicrobial peptide resistance by MprF‐mediated aminoacylation and flipping of phospholipids

Broad‐spectrum antimicrobial peptide resistance by MprF‐mediated aminoacylation and flipping of phospholipids

Lysyl-Phosphatidylglycerol Attenuates Membrane Perturbation Rather than Surface Association of the Cationic Antimicrobial Peptide 6W-RP-1 in a Model Membrane System: Implications for Daptomycin Resistance

Characterization of Two Seryl-tRNA Synthetases in Albomycin-Producing Streptomyces sp. Strain ATCC 700974

Contact Info

Product

Resources

About