Interaction between ATP, oleandomycin and the OleB ATP-binding cassette transporter of <i>Streptomyces antibioticus</i> involved in oleandomycin secretion

Buche, André; Méndez, Cármen; Salas, José A.

doi:10.1042/bj3210139

Cited by 25 publications

(21 citation statements)

References 28 publications

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“…It has been reported that either one of the two ABC domains of Ole(B) is sufficient to confer resistance to oleandomycin (30). Moreover, it was demonstrated that the N-terminal ABC domain of Ole(B) is able to recognize oleandomycin (8). To test the possibility that each ABC domain of Vga(A) would behave similarly in conferring resistance to SGA, we constructed truncated versions of vga(A) by subcloning each half of the gene on separate plasmids.…”

Section: Resultsmentioning

confidence: 99%

Molecular Analysis of Resistance to Streptogramin A Compounds Conferred by the Vga Proteins of Staphylococci

Chesneau¹,

Ligeret²,

Hosan-Aghaie³

et al. 2005

Antimicrob Agents Chemother

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The Vga and Msr resistance determinants, encoded by mobile genetic elements in various staphylococcal strains, belong to a family of ATP-binding cassette (ABC) proteins whose functions and structures are ill defined. Their amino acid sequences are similar to those of proteins involved in the immunity of streptomycetes to the macrolide-lincosamide-streptogramin antibiotics that they produce. Sequence analysis of the genomes of the gram-positive bacteria with low G؉C contents revealed that Lmo0919 from Listeria monocytogenes is more closely related to Vga variants than to Msr variants. In the present study we compared the antibiotic resistance profiles conferred by the Vga-like proteins in two staphylococcal hosts. It was shown that Vga(A), the Vga(A) variant [Vga(A)v], and Lmo0919 can confer resistance to lincosamides and streptogramin A compounds, while only Vga(B) is able to increase the level of resistance to pristinamycin, a mixture of streptogramin A and streptogramin B compounds. By using polyclonal antibodies, we found that the Vga(A) protein colocalized with the ␤ subunit of the F 1 -F 0 ATPase in the membrane fractions of staphylococcal cells. In order to identify functional units in these atypical ABC proteins, such as regions that might be involved in substrate specificity and/or membrane targeting, we analyzed the resistance phenotypes conferred by various plasmids carrying parts or modified versions of the vga(A) gene and we determined the subcellular localization of the gene products. Only polypeptides composed of two ABC domains were detected in the cell membranes. No region of drug specificity was identified. Resistance properties were dependent on the integrities of both Walker B motifs.ATP-binding cassette (ABC) systems share a highly conserved ATP-binding and -hydrolyzing domain or protein. More than 95% of the ABC systems are transporters made up of two hydrophobic transmembrane domains associated with two hydrophilic ATP-binding domains (18). Antibiotic resistance mediated by ABC systems is usually attributable to efflux pumps containing these four core domains (19,21,23,28,31,41). By contrast, a growing number of proteins that confer resistance to macrolide, lincosamide, and streptogramin (MLS) antibiotics consist of a single polypeptide chain carrying two fused hydrophilic ABC domains (for a review, see reference 33). They form a distinct subfamily named ARE (for antibiotic resistance) in the phylogenetic and functional classification of the ABC proteins (6).Many members of the ARE subfamily have been found in MLS producers, such as Car(A) in Streptomyces thermotolerans, Srm(B) in Streptomyces ambofaciens, Tlr(C) in Streptomyces fradiae, Ole(B) in Streptomyces antibioticus, and Lmr(C) in Streptomyces lincolnensis (for a review, see reference 28). Others have been described in pathogens, mainly gram-positive bacteria, where they are either encoded by transposable elements, such as Mel or Orf5 in streptococci (10, 13, 37), or by the chromosome, such as Lsa recently characterized in Enterococc...

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Section: Resultsmentioning

confidence: 99%

Molecular Analysis of Resistance to Streptogramin A Compounds Conferred by the Vga Proteins of Staphylococci

Chesneau¹,

Ligeret²,

Hosan-Aghaie³

et al. 2005

Antimicrob Agents Chemother

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“…Proteins of this subfamily are involved in MLS antibiotic resistance. To the best of our knowledge, only a fragment of Ole(B), a protein belonging to this subfamily, has been produced and purified successfully (24). However, this construct consisted of a fusion of the E. coli maltose-binding protein to the N-terminal ABC domain of Ole(B).…”

Section: Discussionmentioning

confidence: 99%

“…Many ARE subfamily proteins were found in MLS antibiotic-producing actinomycetes (23). The most studied ARE proteins are Ole(B), from the oleandomycin producer Streptomyces antibioticus (24), and two proteins of distinct specificities: Msr(A) and Vga(A), both of which are coded by mobile genetic elements in staphylococci. Msr(A) confers a high level resistance to 14-and 15-membered ring macrolides and to SgB components (25), whereas Vga(A) confers a low level resistance to lincosamides and a high level resistance to SgA components (14).…”

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confidence: 99%

ATP Hydrolysis and Pristinamycin IIA Inhibition of the Staphylococcus aureus Vga(A), a Dual ABC Protein Involved in Streptogramin A Resistance

Jacquet

Girard

Ramaen

et al. 2008

Journal of Biological Chemistry

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In Gram-positive bacteria, a large subfamily of dual ATPbinding cassette proteins confers acquired or intrinsic resistance to macrolide, lincosamide, and streptogramin antibiotics by a far from well understood mechanism. Here, we report the first biochemical characterization of one such protein, Vga(A), which is involved in streptogramin A (SgA) resistance among staphylococci. Vga(A) is composed of two nucleotide-binding domains (NBDs), separated by a charged linker, with a C-terminal extension and without identified transmembrane domains. Highly purified Vga(A) displays a strong ATPase activity (K m ‫؍‬ 78 M, V m ‫؍‬ 6.8 min ؊1 ) that was hardly inhibited by orthovanadate. Using mutants of the conserved catalytic glutamate residues, the two NBDs of Vga(A) were shown to contribute unequally to the total ATPase activity, the mutation at NBD2 being more detrimental than the other. ATPase activity of both catalytic sites was essential for Vga(A) biological function because each single Glu mutant was unable to confer SgA resistance in the staphylococcal host. Of great interest, Vga(A) ATPase was specifically inhibited in a non-competitive manner by the SgA substrate, pristinamycin IIA (PIIA). A deletion of the last 18 amino acids of Vga(A) slightly affected the ATPase activity without modifying the PIIA inhibition values. In contrast, this deletion reduced 4-fold the levels of SgA resistance. Altogether, our results suggest a role for the C terminus in regulation of the SgA antibiotic resistance mechanism conferred by Vga(A) and demonstrate that this dual ATP-binding cassette protein interacts directly and specifically with PIIA, its cognate substrate.

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“…Binding of oleandomycin to a maltose-BP-OleB fusion protein was detected by intrinsic fluorescence (56), but the apparent affinity for the antibiotic was low, in the millimolar range, and no saturation of the signal was reached due to competing absorbance of the compound itself. Given that no interacting TM domains have been identified in this family, substrate specificity may be determined by the ABC proteins.…”

Section: Art Family Of Proteins With Diverse Functionsmentioning

confidence: 99%

Structure, Function, and Evolution of Bacterial ATP-Binding Cassette Systems

Davidson

Dassa

Orelle

et al. 2008

Microbiol Mol Biol Rev

1,187

1,137

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SUMMARY ATP-binding cassette (ABC) systems are universally distributed among living organisms and function in many different aspects of bacterial physiology. ABC transporters are best known for their role in the import of essential nutrients and the export of toxic molecules, but they can also mediate the transport of many other physiological substrates. In a classical transport reaction, two highly conserved ATP-binding domains or subunits couple the binding/hydrolysis of ATP to the translocation of particular substrates across the membrane, through interactions with membrane-spanning domains of the transporter. Variations on this basic theme involve soluble ABC ATP-binding proteins that couple ATP hydrolysis to nontransport processes, such as DNA repair and gene expression regulation. Insights into the structure, function, and mechanism of action of bacterial ABC proteins are reported, based on phylogenetic comparisons as well as classic biochemical and genetic approaches. The availability of an increasing number of high-resolution structures has provided a valuable framework for interpretation of recent studies, and realistic models have been proposed to explain how these fascinating molecular machines use complex dynamic processes to fulfill their numerous biological functions. These advances are also important for elucidating the mechanism of action of eukaryotic ABC proteins, because functional defects in many of them are responsible for severe human inherited diseases.

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Interaction between ATP, oleandomycin and the OleB ATP-binding cassette transporter of Streptomyces antibioticus involved in oleandomycin secretion

Cited by 25 publications

References 28 publications

Molecular Analysis of Resistance to Streptogramin A Compounds Conferred by the Vga Proteins of Staphylococci

Molecular Analysis of Resistance to Streptogramin A Compounds Conferred by the Vga Proteins of Staphylococci

ATP Hydrolysis and Pristinamycin IIA Inhibition of the Staphylococcus aureus Vga(A), a Dual ABC Protein Involved in Streptogramin A Resistance

Structure, Function, and Evolution of Bacterial ATP-Binding Cassette Systems

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