Bacterial resistance to streptothricins

Haupt, Ina; Thrum, H.

doi:10.1002/jobm.3620250508

Cited by 9 publications

(6 citation statements)

References 15 publications

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“…Nourseothricin (NAT) is an inhibitor of ribosomal protein synthesis produced by Streptomyces noursei that induces miscoding during translation in a wide range of prokaryotic and eukaryotic organisms. NAT resistance can be conferred by the S. noursei nat1 gene, encoding an N‐acetyltransferase that monoacetylates nourseothricin (Haupt and Thrum, 1985; Krugel et al , 1993) and has previously been used as a selectable marker in other yeast organisms (Goldstein and McCusker, 1999; McDade and Cox, 2001).…”

Section: Resultsmentioning

confidence: 99%

Three novel antibiotic marker cassettes for gene disruption and marker switching inSchizosaccharomyces pombe

Hentges

Driessche

Tafforeau

et al. 2005

Yeast

259

258

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The ease of construction of multiple mutant strains in Schizosaccharomyces pombe is limited by the number of available genetic markers. We describe here three new cassettes for PCR-mediated gene disruption that can be used in combination with commonly used fission yeast markers to make multiple gene deletions. The natMX6, hphMX6 and bleMX6 markers give rise to resistance towards the antibiotics nourseothricin (NAT), hygromycin B and phleomycin, respectively. The cassettes are composed of exogenous sequences to increase the frequency of integration at targeted loci, and have a structure similar to the commonly used pFA6a-kanMX6 modular plasmid system. This allows a simple exchange of the kanMX6 marker in existing strains with any of the three new cassettes. Alternatively, oligonucleotide primers designed for the modular kanMX6 cassettes can be used to make the transforming PCR fragments for gene disruption. We illustrate the construction of a mutant strain with six independent gene disruptions, using the novel antibiotic cassettes in combination with existing genetic markers.

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Section: Resultsmentioning

confidence: 99%

Three novel antibiotic marker cassettes for gene disruption and marker switching inSchizosaccharomyces pombe

Hentges

Driessche

Tafforeau

et al. 2005

Yeast

259

258

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“…Based on the Tn5-based hyperactive in vitro transposase-based system and broadly expressed proD promoter-driven reporter expression, the toolkit should prove broadly applicable to a wide variety of Gram-negative and -positive bacterial species (38). In addition, the available nonantibiotic selectable markers encoding resistance to nourseothricin (39,40), phleomycin (41), and hygromycin provide multiple options for selection without imparting resistance to therapeutic antibiotics. Such antibiotic resistance when not normally found in pathogens under study might otherwise limit therapeutic options and is potentially proscribed under "dual use research of concern" (42) policies (http://www.phe.gov/s3/dualuse/documents/durc-policy.pdf).…”

Section: Discussionmentioning

confidence: 99%

Promotion and Rescue of Intracellular Brucella neotomae Replication during Coinfection with Legionella pneumophila

Kang

2017

Infect Immun

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We established a new Brucella neotomae in vitro model system for study of type IV secretion system-dependent (T4SS) pathogenesis in the Brucella genus. Importantly, B. neotomae is a rodent pathogen, and unlike B. abortus, B. melitensis, and B. suis, B. neotomae has not been observed to infect humans. It therefore can be handled more facilely using biosafety level 2 practices. More particularly, using a series of novel fluorescent protein and lux operon reporter systems to differentially label pathogens and track intracellular replication, we confirmed T4SS-dependent intracellular growth of B. neotomae in macrophage cell lines. Furthermore, B. neotomae exhibited early endosomal (LAMP-1) and late endoplasmic reticulum (calreticulin)-associated phagosome maturation. These findings recapitulate prior observations for human-pathogenic Brucella spp. In addition, during coinfection experiments with Legionella pneumophila, we found that defective intracellular replication of a B. neotomae T4SS virB4 mutant was rescued and baseline levels of intracellular replication of wild-type B. neotomae were significantly stimulated by coinfection with wild-type but not T4SS mutant L. pneumophila. Using confocal microscopy, it was determined that intracellular colocalization of B. neotomae and L. pneumophila was required for rescue and that colocalization came at a cost to L. pneumophila fitness. These findings were not completely expected based on known temporal and qualitative differences in the intracellular life cycles of these two pathogens. Taken together, we have developed a new system for studying in vitro Brucella pathogenesis and found a remarkable T4SS-dependent interplay between Brucella and Legionella during macrophage coinfection.KEYWORDS Brucella, Legionella pneumophila, coinfection, fluorescent image analysis, intracellular pathogens, pathogenesis, phagosomes, reporter genes, type IV secretion system B rucella species are Gram-negative Alphaproteobacteria that cause chronic, systemic infections in mammals and zoonotic infections in humans (1). These pathogens are known to infect the reticuloendothelial system and proliferate significantly in macrophage-rich organs such as liver, spleen, and bone marrow. Chronic, often debilitating bloodstream infection is typical. In humans, a chronic course punctuated by spikes in body temperature is underscored by the descriptive name, undulant fever, given to disease caused by these organisms. Endovascular infection and osteomyelitis are concerning sequelae. Humans may acquire Brucella from airborne exposure related to large quantities of organisms shed from birthing livestock or from ingesting unpasteurized dairy products.B. abortus, B. melitensis, and B. suis, the species responsible for human zoonotic infection, are facultative intracellular pathogens (1). Intracellular growth is thought critical to successful infection of the host. More particularly, each of these pathogens deploys a type IV secretion system (T4SS), a molecular syringe, to inject virulence

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“…Both streptothricin and nourseothricin are inactivated by acetyltransferase activity [73,92] present in extracts of their producers (Streptomyces strain V-13-1 and Streptomyces noursei, respectively). Genes encoding such activity (sta from Streptomyces lavendulae [80], another streptothricin producer; nat, formerly ntr, from S. noursei [105]) confer high-level resistance in S. lividans and the product generated by purified streptothricin acetyltransferase (STAT) has been characterized as N b -acetylstreptothricin.…”

Section: Streptothricin and Nourseothricinmentioning

confidence: 99%

Avoidance of suicide in antibiotic-producing microbes

Cundliffe

Demain

2010

J Ind Microbiol Biotechnol

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Many microbes synthesize potentially autotoxic antibiotics, mainly as secondary metabolites, against which they need to protect themselves. This is done in various ways, ranging from target-based strategies (i.e. modification of normal drug receptors or de novo synthesis of the latter in drug-resistant form) to the adoption of metabolic shielding and/or efflux strategies that prevent drug-target interactions. These self-defence mechanisms have been studied most intensively in antibiotic-producing prokaryotes, of which the most prolific are the actinomycetes. Only a few documented examples pertain to lower eukaryotes while higher organisms have hardly been addressed in this context. Thus, many plant alkaloids, variously described as herbivore repellents or nitrogen excretion devices, are truly antibiotics-even if toxic to humans. As just one example, bulbs of Narcissus spp. (including the King Alfred daffodil) accumulate narciclasine that binds to the larger subunit of the eukaryotic ribosome and inhibits peptide bond formation. However, ribosomes in the Amaryllidaceae have not been tested for possible resistance to narciclasine and other alkaloids. Clearly, the prevalence of suicide avoidance is likely to extend well beyond the remit of the present article.

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Bacterial resistance to streptothricins

Cited by 9 publications

References 15 publications

Three novel antibiotic marker cassettes for gene disruption and marker switching inSchizosaccharomyces pombe

Three novel antibiotic marker cassettes for gene disruption and marker switching inSchizosaccharomyces pombe

Promotion and Rescue of Intracellular Brucella neotomae Replication during Coinfection with Legionella pneumophila

Avoidance of suicide in antibiotic-producing microbes

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