A spectinomyciin resistance determinant from the spectinomycin producerStreptomyces flavopersicus

Lyutzkanova, D.; Distler, Jürgen; Altenbuchner, Josef

doi:10.1099/00221287-143-7-2135

Cited by 28 publications

(16 citation statements)

References 45 publications

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“…This level of sequence similarity is also observed for CapP with functionally characterized APHs that reside within the biosynthetic gene cluster for a particular aminoglycoside, including the APH from Streptomyces flavopersi-cus that confers resistance to spectinomycin (18/30; accession no. AAB66655) (23).…”

Section: Resultsmentioning

confidence: 99%

Functional and Kinetic Analysis of the Phosphotransferase CapP Conferring Selective Self-resistance to Capuramycin Antibiotics

Yang

Funabashi

Nonaka

et al. 2010

Journal of Biological Chemistry

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Capuramycin-related compounds, including A-500359s and A-503083s, are nucleoside antibiotics that inhibit the enzyme bacterial translocase I involved in peptidoglycan cell wall biosynthesis. Within the biosynthetic gene cluster for the A-500359s exists a gene encoding a putative aminoglycoside 3-phosphotransferase that was previously demonstrated to be highly expressed during the production of A-500359s and confers selective resistance to capuramycins when expressed in heterologous hosts. A similar gene (capP) was identified within the biosynthetic gene cluster for the A-503083s, and CapP is now shown to similarly confer selective resistance to capuramycins. Recombinant CapP was produced and purified from Escherichia coli, and the function of CapP is established as an ATP-dependent capuramycin phosphotransferase that regio-specifically transfers the ␥-phosphate to the 3؆-hydroxyl of the unsaturated hexuronic acid moiety of A-503083 B. Kinetic analysis with the three major A-503083 congeners suggests that CapP preferentially phosphorylates A-503083s containing an aminocaprolactam moiety attached to the hexuronic acid, and bi-substrate kinetic analysis was consistent with CapP employing a sequential kinetic mechanism similar to most known aminoglycoside 3-phosphotransferases. The purified CapP product lost its antibiotic activity against Mycobacterium smegmatis, and this loss in bioactivity is primarily due to a 272-fold increase in the IC 50 in the bacterial translocase I-catalyzed reaction. The results establish CapP-mediated phosphorylation as a mechanism of resistance to capuramycins and now set the stage to explore this strategy of resistance as a potential mechanism inherent to pathogens and provide the impetus for preparing second generation analogues as a preemptive strike to such resistance strategies.Infectious and parasitic diseases are the second leading cause of death worldwide, and this includes more than 1.6 million deaths by tuberculosis (TB) 4 in 2006 (1). Alarmingly, nearly 5% of all of the new cases of TB during this same year were caused by Mycobacterium tuberculosis, the primary causative agent of TB, that had resistance to at least one of the commonly used antibiotics to treat TB (2). To compound this problematic emergence of drug resistance, the discovery and development of new antibiotics have continued to decline with only a few new classes of antibiotics introduced into the clinic within the past few decades (3, 4). Thus, there is a great need for new antibiotics with novel modes of action and unique structures to combat multiple drug-resistant pathogens such as M. tuberculosis.Capuramycin was discovered from Streptomyces griseus 446-S3 based on gross antibacterial activity (5), and analogues termed A-500359s from S. griseus SANK 60196 (6 -8) and A-503083s from Streptomyces sp. SANK 62799 (9) were subsequently discovered using a specific screen aimed at identifying bacterial translocase I (MraY) inhibitors. MraY initiates the lipid cycle of peptidoglycan cell wall biosynthesis, a pathway ...

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Section: Resultsmentioning

confidence: 99%

Functional and Kinetic Analysis of the Phosphotransferase CapP Conferring Selective Self-resistance to Capuramycin Antibiotics

Yang

Funabashi

Nonaka

et al. 2010

Journal of Biological Chemistry

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“…MppL was similar to the spectinomycin resistance pump SpcT from Streptomyces flavopersicus (a spectinomycin producer) and the putative viomycin self-resistance determinant VioE from Streptomyces sp. strain ATCC 11861 (25,38). To determine the role of MppL, a strain (WNP103) carrying a null form of mppL [mppL::aac(3)IV] was constructed.…”

Section: Resultsmentioning

confidence: 99%

Biosynthetic Pathway for Mannopeptimycins, Lipoglycopeptide Antibiotics Active against Drug-Resistant Gram-Positive Pathogens

Magarvey¹,

Haltli²,

He³

et al. 2006

Antimicrob Agents Chemother

110

116

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The mannopeptimycins are a novel class of lipoglycopeptide antibiotics active against multidrug-resistant pathogens with potential as clinically useful antibacterials. This report is the first to describe the biosynthesis of this novel class of mannosylated lipoglycopeptides. Included here are the cloning, sequencing, annotation, and manipulation of the mannopeptimycin biosynthetic gene cluster from Streptomyces hygroscopicus NRRL 30439. Encoded by genes within the mannopeptimycin biosynthetic gene cluster are enzymes responsible for the generation of the hexapeptide core (nonribosomal peptide synthetases [NRPS]) and tailoring reactions (mannosylation, isovalerylation, hydroxylation, and methylation). The NRPS system is noncanonical in that it has six modules utilizing only five amino acid-specific adenylation domains and it lacks a prototypical NRPS macrocyclizing thioesterase domain. Analysis of the mannopeptimycin gene cluster and its engineering has elucidated the mannopeptimycin biosynthetic pathway and provides the framework to make new and improved mannopeptimycins biosynthetically.Multidrug-resistant gram-positive bacterial pathogens now exist, and their rising numbers are a major concern. The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) exemplifies the seriousness of this concern (22, 29). Finding novel antibiotics active against such drug-resistant gram-positive pathogens, however, is problematic (32, 42). The mannopeptimycins ( Fig. 1), which are produced by Streptomyces hygroscopicus NRRL 30439, represent a new class of lipoglycopeptide antibiotics with exceptional in vitro and in vivo antibacterial activities against MRSA, VRE, and penicillin-resistant Streptococcus pneumoniae (17,36). Studies with mannopeptimycin-␦ demonstrated that this antibiotic acts by blocking the transglycosylation reaction in cell wall biosynthesis mediated by lipid II binding. Mannopeptimycin-␦ inhibits gram-positive cell wall biosynthesis through a mechanism which does not compete or render it ineffective due to crossresistance with the vancomycin-type antibiotics (33, 36).The unique structure, mode of action, and bioactivity of the mannopeptimycins have generated much interest in identifying a therapeutic candidate from this new class of molecules (17,33,36). One semisynthetic analog, AC98-6446, of the natural mannopeptimycins is significantly more potent and effective than the natural mannopeptimycins, with MICs in the 15-to 60-ng/ml range against MRSA, VRE, penicillin-resistant Streptococcus pneumoniae, and glycopeptide-intermediate Staphylococcus aureus (10, 31). The success in improving on the natural mannopeptimycins argues strongly for broadening the effort of generating additional new mannopeptimycins through promising strategies such as combinatorial biosynthesis, mutasynthesis, and chemoenzymatic synthesis (4, 28). The establishment and success of such approaches hinge on the cloning of the mannopeptimycin biosynthetic gene cluster and elucidation of ...

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“…Aminoglycoside phosphotransferases APH(4) and APH(7Љ) produce resistance to hygromycin (108,324), while APH(9) produces resistance to spectinomycin (171,282). These enzymes do not present any clinical significance and will not be discussed here in detail.…”

Section: Aminoglycoside Phosphotransferasesmentioning

confidence: 99%

Versatility of Aminoglycosides and Prospects for Their Future

2003

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Aminoglycoside antibiotics have had a major impact on our ability to treat bacterial infections for the past half century. Whereas the interest in these versatile antibiotics continues to be high, their clinical utility has been compromised by widespread instances of resistance. The multitude of mechanisms of resistance is disconcerting but also illuminates how nature can manifest resistance when bacteria are confronted by antibiotics. This article reviews the most recent knowledge about the mechanisms of aminoglycoside action and the mechanisms of resistance to these antibiotics

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A spectinomyciin resistance determinant from the spectinomycin producerStreptomyces flavopersicus

Cited by 28 publications

References 45 publications

Functional and Kinetic Analysis of the Phosphotransferase CapP Conferring Selective Self-resistance to Capuramycin Antibiotics

Functional and Kinetic Analysis of the Phosphotransferase CapP Conferring Selective Self-resistance to Capuramycin Antibiotics

Biosynthetic Pathway for Mannopeptimycins, Lipoglycopeptide Antibiotics Active against Drug-Resistant Gram-Positive Pathogens

Versatility of Aminoglycosides and Prospects for Their Future

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