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Macrolide phosphotransferase

Corey Fyfe, Trudy H Grossman, Kathy Kerstein, Joyce Sutcliffe
Macrolide resistance mechanisms can be target-based with a change in a 23S ribosomal RNA (rRNA) residue or a mutation in ribosomal protein L4 or L22 affecting the ribosome's interaction with the antibiotic. Alternatively, mono- or dimethylation of A2058 in domain V of the 23S rRNA by an acquired rRNA methyltransferase, the product of an erm (erythromycin ribosome methylation) gene, can interfere with antibiotic binding. Acquired genes encoding efflux pumps, most predominantly mef(A) + msr(D) in pneumococci/streptococci and msr(A/B) in staphylococci, also mediate resistance...
October 3, 2016: Cold Spring Harbor Perspectives in Medicine
Longping Li, Weiwei Feng, Zhiping Zhang, Huping Xue, Xin Zhao
BACKGROUND: There are limited data available on macrolide-lincosamide-streptogramin (MLS) resistance of Staphylococcus aureus (S. aureus) and coagulase-negative staphylococci (CoNS) from bovine milk in China. To address this knowledge gap, MLS resistance was determined in 121 S. aureus and 97 CoNS isolates. Minimum inhibitory concentrations (MICs) of MLS antibiotics were determined by an agar dilution method, while differentiation of MLS phenotypes was performed by a double-disc diffusion test...
2015: BMC Veterinary Research
L Nonaka, F Maruyama, S Suzuki, M Masuda
UNLABELLED: The aim of this study was to determine whether mef(C) and mph(G), originally found on the transferable multi-drug plasmid pAQU1 from Photobacterium damselae subsp. damselae isolated from seawater of a fish farm, are responsible for conferring macrolide resistance. Since these genes are localized head-to-tail on pAQU1 and only four nucleotides exist between them, the single- and combination-effect of these genes was examined. When mph(G) alone was introduced to Escherichia coli, the minimum inhibitory concentrations (MICs) against erythromycin, clarithromycin and azithromycin increased, whereas introduction of mef(C) alone did not influence macrolide susceptibility...
July 2015: Letters in Applied Microbiology
Chao Wang, Zhihai Sui, Sébastien Olivier Leclercq, Gang Zhang, Meilin Zhao, Weiqi Chen, Jie Feng
The Bacillus cereus group is composed of Gram-positive spore-forming bacteria of clinical and ecological importance. More than 200 B. cereus group isolates have been sequenced. However, there are few reports of B. cereus group antibiotic resistance genes. This study identified two functional classes of macrolide phosphotransferases (Mphs) in the B. cereus group. Cluster A Mphs inactivate 14- and 15-membered macrolides while Cluster B Mphs inactivate 14-, 15- and 16-membered compounds. The genomic region surrounding the Cluster B Mph gene is related to various plasmid sequences, suggesting that this gene is an acquired resistance gene...
May 2015: Environmental Microbiology
Jing Yang, Chao Wang, Jinyu Wu, Li Liu, Gang Zhang, Jie Feng
The genus Exiguobacterium can adapt readily to, and survive in, diverse environments. Our study demonstrated that Exiguobacterium sp. strain S3-2, isolated from marine sediment, is resistant to five antibiotics. The plasmid pMC1 in this strain carries seven putative resistance genes. We functionally characterized these resistance genes in Escherichia coli, and genes encoding dihydrofolate reductase and macrolide phosphotransferase were considered novel resistance genes based on their low similarities to known resistance genes...
February 2014: Applied and Environmental Microbiology
Nicolas Desroy, Alexis Denis, Chrystelle Oliveira, Dmytro Atamanyuk, Sophia Briet, Fabien Faivre, Géraldine LeFralliec, Yannick Bonvin, Mayalen Oxoby, Sonia Escaich, Stéphanie Floquet, Elodie Drocourt, Vanida Vongsouthi, Lionel Durant, François Moreau, Theodore B Verhey, Ting-Wai Lee, Murray S Junop, Vincent Gerusz
We report here the optimization of an HldE kinase inhibitor to low nanomolar potency, which resulted in the identification of the first reported compounds active on selected E. coli strains. One of the most interesting candidates, compound 86, was shown to inhibit specifically bacterial LPS heptosylation on efflux pump deleted E. coli strains. This compound did not interfere with E. coli bacterial growth (MIC > 32 μg/mL) but sensitized this pathogen to hydrophobic antibiotics like macrolides normally inactive on Gram-negative bacteria...
February 28, 2013: Journal of Medicinal Chemistry
Simon Rose, Benoit Desmolaize, Puneet Jaju, Cornelia Wilhelm, Ralf Warrass, Stephen Douthwaite
The bacterial pathogens Mannheimia haemolytica and Pasteurella multocida are major etiological agents in respiratory tract infections of cattle. Although these infections can generally be successfully treated with veterinary macrolide antibiotics, a few recent isolates have shown resistance to these drugs. Macrolide resistance in members of the family Pasteurellaceae is conferred by combinations of at least three genes: erm(42), which encodes a monomethyltransferase and confers a type I MLS(B) (macrolide, lincosamide, and streptogramin B) phenotype; msr(E), which encodes a macrolide efflux pump; and mph(E), which encodes a macrolide-inactivating phosphotransferase...
July 2012: Antimicrobial Agents and Chemotherapy
Kirandeep Bhullar, Nicholas Waglechner, Andrew Pawlowski, Kalinka Koteva, Eric D Banks, Michael D Johnston, Hazel A Barton, Gerard D Wright
Antibiotic resistance is a global challenge that impacts all pharmaceutically used antibiotics. The origin of the genes associated with this resistance is of significant importance to our understanding of the evolution and dissemination of antibiotic resistance in pathogens. A growing body of evidence implicates environmental organisms as reservoirs of these resistance genes; however, the role of anthropogenic use of antibiotics in the emergence of these genes is controversial. We report a screen of a sample of the culturable microbiome of Lechuguilla Cave, New Mexico, in a region of the cave that has been isolated for over 4 million years...
2012: PloS One
Janet N Y Chan, Dajana Vuckovic, Lekha Sleno, Jonathan B Olsen, Oxana Pogoutse, Pierre Havugimana, Johannes A Hewel, Navgeet Bajaj, Yale Wang, Marcel F Musteata, Corey Nislow, Andrew Emili
Bioactive molecules typically mediate their biological effects through direct physical association with one or more cellular proteins. The detection of drug-target interactions is therefore essential for the characterization of compound mechanism of action and off-target effects, but generic label-free approaches for detecting binding events in biological mixtures have remained elusive. Here, we report a method termed target identification by chromatographic co-elution (TICC) for routinely monitoring the interaction of drugs with cellular proteins under nearly physiological conditions in vitro based on simple liquid chromatographic separations of cell-free lysates...
July 2012: Molecular & Cellular Proteomics: MCP
Laura Gardner, Yan Zou, Alexandria Mara, T Ashton Cropp, Alexander Deiters
Bacterial cells control resistance to the macrolide antibiotic erythromycin using the MphR(A) repressor protein. Erythromycin binds to MphR(A), causing release of the PmphR promoter, activating expression of the 2'-phosphotransferase Mph(A). We engineered the MphR(A)/promoter system to, in conjunction with a light-activatable derivative of erythromycin, enable photochemical activation of gene expression in E. coli. We applied this photochemical gene switch to the construction of a light-triggered logic gate, a light-controlled band-pass filter, as well as spatial and temporal control of gene expression...
September 2011: Molecular BioSystems
Benoit Desmolaize, Simon Rose, Cornelia Wilhelm, Ralf Warrass, Stephen Douthwaite
Respiratory tract infections in cattle are commonly associated with the bacterial pathogens Mannheimia haemolytica and Pasteurella multocida. These infections can generally be successfully treated in the field with one of several groups of antibiotics, including macrolides. A few recent isolates of these species exhibit resistance to veterinary macrolides with phenotypes that fall into three distinct classes. The first class has type I macrolide, lincosamide, and streptogramin B antibiotic resistance and, consistent with this, the 23S rRNA nucleotide A2058 is monomethylated by the enzyme product of the erm(42) gene...
September 2011: Antimicrobial Agents and Chemotherapy
Kristina Kadlec, Geovana Brenner Michael, Michael T Sweeney, Elzbieta Brzuszkiewicz, Heiko Liesegang, Rolf Daniel, Jeffrey L Watts, Stefan Schwarz
The mechanism of macrolide-triamilide resistance in Pasteurella multocida has been unknown. During whole-genome sequencing of a multiresistant bovine P. multocida isolate, three new resistance genes, the rRNA methylase gene erm(42), the macrolide transporter gene msr(E), and the macrolide phosphotransferase gene mph(E), were detected. The three genes were PCR amplified, cloned into suitable plasmid vectors, and shown to confer either macrolide-lincosamide resistance [erm(42)] or macrolide-triamilide resistance [msr(E)-mph(E)] in macrolide-susceptible Escherichia coli and P...
May 2011: Antimicrobial Agents and Chemotherapy
Rebecca Leigh Howie, Jason P Folster, Anna Bowen, Ezra J Barzilay, Jean M Whichard
To investigate azithromycin susceptibility in Shigella sonnei in the United States, we examined the azithromycin minimum inhibitory concentrations (MICs) of outbreak and routine human S. sonnei isolates. Isolate susceptibility clustered at 8 mg/L, but three isolates displayed higher MICs (>64  mg/L) to azithromycin. All three isolates contained a plasmid-encoded mphA gene, known to encode a macrolide-2'-phosphotransferase enzyme. Transformation of the mphA gene into Escherichia coli DH10B allowed the transfer of decreased susceptibility to azithromycin...
December 2010: Microbial Drug Resistance: MDR: Mechanisms, Epidemiology, and Disease
Tushar Shakya, Gerard D Wright
Antibiotic kinases, which include aminoglycoside and macrolide phosphotransferases (APHs and MPHs), pose a serious threat to currently used antimicrobial therapies. These enzymes show structural and functional homology with Ser/Thr/Tyr kinases, which is suggestive of a common ancestor. Surprisingly, recent in vitro studies using purified antibiotic kinase enzymes have revealed that a number are able to utilize GTP as the antibiotic phospho donor, either preferentially or exclusively compared to ATP, the canonical phosphate donor in most biochemical reactions...
May 2010: Antimicrobial Agents and Chemotherapy
Jianting Zheng, Vatsala Sagar, Adam Smolinsky, Chase Bourke, Nicole LaRonde-LeBlanc, T Ashton Cropp
The regulatory protein MphR(A) has recently seen extensive use in synthetic biological applications, such as metabolite sensing and exogenous control of gene expression. This protein negatively regulates the expression of a macrolide 2'-phosphotransferase I resistance gene (mphA) via binding to a 35-bp DNA operator upstream of the start codon and is de-repressed by the presence of erythromycin. Here, we present the refined crystal structure of the MphR(A) protein free of erythromycin and that of the MphR(A) protein with bound erythromycin at 2...
April 17, 2009: Journal of Molecular Biology
Leyla Boumghar-Bourtchai, Patricia Mariani-Kurkdjian, Edouard Bingen, Ingrid Filliol, Anne Dhalluin, Shadia Ait Ifrane, François-Xavier Weill, Roland Leclercq
Shigella sonnei UCN59, isolated during an outbreak of S. sonnei in January 2007, was resistant to azithromycin (MIC 64 mg/L). The isolate contained a plasmid-borne mph(A) gene encoding a macrolide 2 -phosphotransferase that inactivates macrolides. Emergence of the mph(A) gene in S. sonnei may limit usefulness of azithromycin for treatment of shigellosis.
August 2008: Emerging Infectious Diseases
Mohammad Emaneini, Marzieh Aligholi, Maneli Aminshahi
The prevalence of glycopeptides, aminoglycosides and erythromycin resistance among Enterococcus faecalis and Enterococcus faecium was investigated. The susceptibility of 326 enterococcal hospital isolates to amikacin, kanamycin, netilmicin and tobramycin were determined using disk diffusion method. The minimum inhibitory concentration (MIC) of vancomycin, teicoplanin, gentamicin, streptomycin, and erythromycin were determined by microbroth dilution method. The genes encoding aminoglycoside modifying enzymes described as AMEs genes, erythromycin-resistant methylase (erm) and vancomycin-resistant were targeted by multiplex-PCR reaction...
2008: Polish Journal of Microbiology
Adeline Achard, Véronique Guérin-Faublée, Vianney Pichereau, Corinne Villers, Roland Leclercq
Streptococcus uberis UCN60 was resistant to spiramycin (MIC = 8 microg/ml) but susceptible to erythromycin (MIC = 0.06 microg/ml), azithromycin (MIC = 0.12 microg/ml), josamycin (MIC = 0.25 microg/ml), and tylosin (MIC = 0.5 microg/ml). A 2.5-kb HindIII fragment was cloned from S. uberis UCN60 DNA on plasmid pUC18 and introduced into Escherichia coli AG100A, where it conferred resistance to spiramycin by inactivation. The sequence analysis of the fragment showed the presence of an rdmC-like gene that putatively encoded a protein belonging to the alpha/beta hydrolase family and of the first 196 nucleotides of the mph(B) gene putatively encoding a phosphotransferase known to inactivate 14-, 15-, and 16-membered macrolides in E...
August 2008: Antimicrobial Agents and Chemotherapy
Romina Camilli, Maria Del Grosso, Francesco Iannelli, Annalisa Pantosti
erm(A) subclass erm(TR), a common macrolide resistance determinant in Streptococcus pyogenes but quite rare in Streptococcus pneumoniae, was found in a clinical S. pneumoniae isolate (AP200) from Italy. In this isolate, erm(TR) was found included in a genetic element approximately 56 kb in size that did not appear to be conjugative but could be transferred by transformation. An erm(TR)-containing DNA fragment of approximately 10 kb was sequenced and 12 open reading frames (ORFs) were identified. Upstream of erm(TR), a regulatory protein of the TetR family and the two components of an efflux pump of the ABC type were found...
February 2008: Antimicrobial Agents and Chemotherapy
Olivier Chesneau, Krassimira Tsvetkova, Patrice Courvalin
This study aims to compare the resistance phenotypes conferred by various genes encoding enzymes that phosphorylate erythromycin. The mph genes were cloned into Escherichia coli AG100A susceptible to macrolides and ketolides following disruption of the AcrAB pump. An 882 bp sequence containing a premature stop codon, homologous to the three other previously described mph genes and present widely among Enterobacteriaceae, was found to confer resistance to erythromycin by phosphorylation. The mph(C) gene, as reported for mph(B), also conferred resistance to spiramycin...
April 2007: FEMS Microbiology Letters
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