An ATP-binding cassette transporter and two rRNA methyltransferases are involved in resistance to avilamycin in the producer organism Streptomyces viridochromogenes Tü57

Three different resistance factors from the avilamycin biosynthetic gene cluster of Streptomyces viridochromogenes Tü57, which confer avilamycin resistance when expressed in Streptomyces lividans TK66, were isolated. Analysis of the deduced amino acid sequences showed that AviABC1 is similar to a la...

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Bibliographic Details
Published in:Antimicrobial agents and chemotherapy 2001-03, Vol.45 (3), p.690-695
Main Authors: WEITNAUER, Gabriele, GAISSER, Sibylle, TREFZER, Axel, STOCKERT, Sigrid, WESTRICH, Lucy, QUIROS, Luis M, MENDEZ, Carmen, SALAS, Jose A, BECHTHOLD, Andreas
Format: Article
Language:English
Subjects:
Anti-Bacterial Agents - pharmacology
ATP-Binding Cassette Transporters - genetics
ATP-Binding Cassette Transporters - metabolism
ATP-binding protein
avi gene
Avilamycins
Bacterial Proteins - genetics
Biological and medical sciences
Chromatography, Affinity
Cloning, Molecular
Drug Resistance, Microbial - genetics
Drug Resistance, Microbial - physiology
Escherichia coli
Fundamental and applied biological sciences. Psychology
Growth, nutrition, metabolism, transports, enzymes. Molecular biology
Mechanisms of Resistance
Methyltransferases - genetics
Methyltransferases - metabolism
Microbial Sensitivity Tests
Microbiology
Molecular Sequence Data
Mycology
Oligosaccharides - pharmacology
Ribosomes - drug effects
rRNA methyltransferase
Streptomyces - drug effects
Streptomyces - metabolism
Streptomyces viridochromogenes
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Summary:Three different resistance factors from the avilamycin biosynthetic gene cluster of Streptomyces viridochromogenes Tü57, which confer avilamycin resistance when expressed in Streptomyces lividans TK66, were isolated. Analysis of the deduced amino acid sequences showed that AviABC1 is similar to a large family of ATP-binding transporter proteins and that AviABC2 resembles hydrophobic transmembrane proteins known to act jointly with the ATP-binding proteins. The deduced amino acid sequence of aviRb showed similarity to those of other rRNA methyltransferases, and AviRa did not resemble any protein in the databases. Independent expression in S. lividans TK66 of aviABC1 plus aviABC2, aviRa, or aviRb conferred different levels of resistance to avilamycin: 5, 10, or 250 microg/ml, respectively. When either aviRa plus aviRb or aviRa plus aviRb plus aviABC1 plus aviABC2 was coexpressed in S. lividans TK66, avilamycin resistance levels reached more than 250 microg/ml. Avilamycin A inhibited poly(U)-directed polyphenylalanine synthesis in an in vitro system using ribosomes of S. lividans TK66(pUWL201) (GWO), S. lividans TK66(pUWL201-Ra) (GWRa), or S. lividans TK66(pUWL201-Rb) (GWRb), whereas ribosomes of S. lividans TK66 containing pUWL201-Ra+Rb (GWRaRb) were highly resistant. aviRa and aviRb were expressed in Escherichia coli, and both enzymes were purified as fusion proteins to near homogeneity. Both enzymes showed rRNA methyltransferase activity using a mixture of 16S and 23S rRNAs from E. coli as the substrate. Coincubation experiments revealed that the enzymes methylate different positions of rRNA.
ISSN:0066-4804
1098-6596
DOI:10.1128/AAC.45.3.690-695.2001