Cloning and characterization of gentamicin-resistance genes from Micromonospora purpurea and Micromonospora rosea

Kelemen, Gabriella H.; Cundliffe, Eric; Financsek, I.

doi:10.1016/0378-1119(91)90103-i

Cited by 43 publications

(20 citation statements)

References 16 publications

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“…Identities with other 16S rRNA methylases produced by Streptomyces and Micromonospora species were generally lower. Amino acid identities of RmtB were 33 and 32% with GrmB and Sgm methylases of sisomicin-producing Micromonospora rosea and Micromonospora zionensis, respectively (14); 32% with GrmA methylase of gentamicin-producing Micromonospora purpurea (14); 31% with Kmr methylase of kanamycin-producing Streptomyces kanamyceticus (8); 30% with FmrO methylase of fortimicin-producing Micromonospora olivasterospora (18); and 27% with KgmB of nebramycin-producing Streptomyces tenebrarius (12). The putative promoter region of rmtB appeared to be located within the right-hand end of Tn3, just upstream of the inverted repeat (Fig.…”

Section: Resultsmentioning

confidence: 99%

Plasmid-Mediated 16S rRNA Methylase in Serratia marcescens Conferring High-Level Resistance to Aminoglycosides

Doi

Yokoyama

Yamane

et al. 2004

Antimicrob Agents Chemother

168

118

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Serratia marcescens S-95, which displayed an unusually high degree of resistance to aminoglycosides, including kanamycins and gentamicins, was isolated in 2002 from a patient in Japan. The resistance was mediated by a large plasmid which was nonconjugative but transferable to an Escherichia coli recipient by transformation. The gene responsible for the aminoglycoside resistance was cloned and sequenced. The deduced amino acid sequence of the resistance gene shared 82% identity with RmtA, which was recently identified as 16S rRNA methylase conferring high-level aminoglycoside resistance in Pseudomonas aeruginosa. Histidine-tagged recombinant protein showed methylation activity against E. coli 16S rRNA. The novel aminoglycoside resistance gene was therefore designated rmtB. The genetic environment of rmtB was further investigated. The sequence immediately upstream of rmtB contained the right end of transposon Tn3, including bla TEM , while an open reading frame possibly encoding a transposase was identified downstream of the gene. This is the first report describing 16S rRNA methylase production in S. marcescens. The aminoglycoside resistance mechanism mediated by production of 16S rRNA methylase and subsequent ribosomal protection used to be confined to aminoglycoside-producing actinomycetes. However, it is now identified among pathogenic bacteria, including Enterobacteriaceae and P. aeruginosa in Japan. This is a cause for concern since other treatment options are often limited in patients requiring highly potent aminoglycosides such as amikacin and tobramycin.

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Section: Resultsmentioning

confidence: 99%

Plasmid-Mediated 16S rRNA Methylase in Serratia marcescens Conferring High-Level Resistance to Aminoglycosides

Doi

Yokoyama

Yamane

et al. 2004

Antimicrob Agents Chemother

168

118

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“…RNA samples were isolated at different time points from plate-grown cultures. The hrdB transcript was used as a positive internal control because hrdB transcription has been reported to be relatively constant throughout development (13). S1 nuclease protection assays with the eshA-specific probe (see Fig.…”

Section: Resultsmentioning

confidence: 99%

Molecular and Functional Analyses of the Gene ( eshA ) Encoding the 52-Kilodalton Protein of Streptomyces coelicolor A3(2) Required for Antibiotic Production

et al. 2001

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Analysis of proteins recovered in the S100 precipitate fraction of Streptomyces griseus after ultracentrifugation led to the identification of a 52-kDa protein which is produced during the late growth phase. The gene (eshA) which codes for this protein was cloned from S. griseus, and then its homologue was cloned from Streptomyces coelicolor A3(2). The protein was deduced to be 471 amino acids in length. The protein EshA is characterized by a central region that shows homology to the eukaryotic-type cyclic nucleotide-binding domains. Significant homology was also found to MMPI in Mycobacterium leprae, a major antigenic protein to humans. The eshA gene mapped near the chromosome end and was not essential for viability, as demonstrated by gene disruption experiments, but its disruption resulted in the abolishment of an antibiotic (actinorhodin but not undecylprodigiosin) production. Aerial mycelium was produced as abundantly as by the parent strain. Expression analysis of the EshA protein by Western blotting revealed that EshA is present only in late-growthphase cells. The eshA gene was transcribed just preceding intracellular accumulation of the EshA protein, as determined by S1 nuclease protection, indicating that EshA expression is regulated at the transcription level. The expression of EshA was unaffected by introduction of the relA mutation, which blocks ppGpp synthesis.Streptomycetes are gram-positive filamentous soil bacteria which produce a wide variety of secondary metabolites that include about half of the known microbial antibiotics. In addition to antibiotic production (physiological differentiation), the genus Streptomyces is also characterized by the ability to form aerial mycelium from vegetative mycelium when grown in solid culture (morphological differentiation). Streptomyces coelicolor A3(2), the most fully genetically characterized streptomycete, is an appropriate strain for studying the regulation of morphological and physiological differentiation (for a review, see the work by Chater and Hopwood [2]). This strain produces at least four antibiotics, of which the blue-pigmented polyketide antibiotic actinorhodin and the red-pigmented antibiotic undecylprodigiosin are usually produced in stationaryphase cultures (reviewed by Chater and Bibb [3]). Much progress has been made in elucidating not only the organization of antibiotic biosynthesis gene clusters in several Streptomyces species but also a number of pathway-specific regulatory genes that are required for the activation of their cognate biosynthesis genes (reviewed by Hunter and Baumberg [7]). In addition to pathway-specific regulatory genes, S. coelicolor possesses several genes that have pleiotropic effects on antibiotic production. These genes fall into two classes: those that affect only antibiotic production (absA, absB, afsB, afsR, and abaA) and those that affect both antibiotic production and morphological differentiation (bldA, bldB, bldC, bldD, bldF, bldG, bldH, bldI, bldJ, bldK, bldL, bldM, and bldN) (for reviews, see reference...

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“…The signals detected in Micromonospora strains should be resistance genes similar to f m r 0 considering the high resistance of these strains to FTM-A. The resistance genes with resistance patterns similar to fmr0 isolated from S. kanamyceticus (Nakano et al, 1984), S. tenebrarius (kgmB) (Skeggs et al, 1987) and M. purpurea (grm) (Kelemen et al, 1991) confer ribosome modification. Ribosomal resistance is a widespread mechanism among aminoglycoside-producing Micromonospora strains (Matkovic et al, 1984).…”

Section: Discussionmentioning

confidence: 97%

“…It was reported that grrn of M . purpurea encoded a specific methyltransferase of 16s ribosomal RNA (Kelemen et al, 1991). However, the size of the band that hybridized to the fmr0 probe in M. purpurea ATCC 15835 (identical to strain DSM 43036 used by Piendl et al, 1984) (6.1 kb BamHI fragment; Fig.…”

Section: Discussionmentioning

confidence: 98%

See 1 more Smart Citation

Characterization of Two Different Types of Resistance Genes Among Producers of Fortimicin-Group Antibiotics

Ohta

Dairi²,

Hasegawa³

1993

Journal of General Microbiology

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Cloning and characterization of gentamicin-resistance genes from Micromonospora purpurea and Micromonospora rosea

Cited by 43 publications

References 16 publications

Plasmid-Mediated 16S rRNA Methylase in Serratia marcescens Conferring High-Level Resistance to Aminoglycosides

Plasmid-Mediated 16S rRNA Methylase in Serratia marcescens Conferring High-Level Resistance to Aminoglycosides

Molecular and Functional Analyses of the Gene ( eshA ) Encoding the 52-Kilodalton Protein of Streptomyces coelicolor A3(2) Required for Antibiotic Production

Characterization of Two Different Types of Resistance Genes Among Producers of Fortimicin-Group Antibiotics

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