Cloning and characterization of two genes from Streptomyces lividans that confer inducible resistance to lincomycin and macrolide antibiotics

Jenkins, G.N.; Cundliffe, Eric

doi:10.1016/0378-1119(91)90487-v

Cited by 84 publications

(74 citation statements)

References 19 publications

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“…The deduced product of srmA is a protein of 259 aa (molecular mass 28n6 kDa) and sequence comparisons with databases showed end-to-end similarity to several rRNA methyltransferases that confer resistance to MLS antibiotics. For instance, SrmA presents 86 % and 75 % identity with the Lrm protein (Jenkins & Cundliffe, 1991) and ErmSV (Kamimiya & Weisblum, 1997), respectively. Also, SrmA contains, between residues 40 and 48, a glycine-rich sequence that is shared by other SAM-dependent methyltransferases (Kagan & Clarke, 1994) and has been shown by crystallographic analysis to be part of the SAM-binding site (Schluckebier et al, 1995 .…”

Section: Nucleotide Sequence Of Srmamentioning

confidence: 99%

Dispensable ribosomal resistance to spiramycin conferred by srmA in the spiramycin producer Streptomyces ambofaciens The EMBL/GenBank accession number for the nucleotide sequence described in this paper is AJ223970.

et al. 1999

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Streptomyces ambofaciens produces the macrolide antibiotic spiramycin, an inhibitor of protein synthesis, and possesses multiple resistance mechanisms to the produced antibiotic. Several resistance determinants have been isolated from S. ambofaciens and studies with one of them, srmA, which hybridized with ermE (the erythromycin-resistance gene from Saccharopolyspora erythraea), are detailed here. The nucleotide sequence of srmA was determined and the mechanism by which its product confers resistance was characterized. The SrmA protein is a methyltransferase which introduces a single methyl group into A-2058 (Escherichia coli numbering scheme) in the large rRNA, thereby conferring an MLS (macrolide-lincosamide-streptogramin type B) type I resistance phenotype. A mutant of S. ambofaciens in which srmA was inactivated was viable and still produced spiramycin, indicating that srmA is dispensable, at least in the presence of the other resistance determinants.

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Section: Nucleotide Sequence Of Srmamentioning

confidence: 99%

Dispensable ribosomal resistance to spiramycin conferred by srmA in the spiramycin producer Streptomyces ambofaciens The EMBL/GenBank accession number for the nucleotide sequence described in this paper is AJ223970.

et al. 1999

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“…The molecular basis for resistance in these strains was subsequently shown by Ross et al (94) to involve the active efflux of erythromycin and streptogramin B but not clindamycin. Additional mechanisms of macrolide resistance, all associated with the acquisition of new genetic information, including structural modification of erythromycin by phosphorylation (82), glycosylation (60), and lactone ring cleavage by erythromycin esterase (2,83), have been added to the list.…”

Section: Introductionmentioning

confidence: 99%

Erythromycin resistance by ribosome modification

Weisblum

1995

Antimicrob Agents Chemother

925

752

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“…Computer analysis of the deduced protein product of dnrH with the BLASTP and TBLASTN programs (1, 9) identified two Streptomyces glycosyl transferases: Mgt, a macrolide glycosyl transferase (17), and OleD, an oleandomycin glycosyl transferase (15). Similarity was also found with a subunit of a rhamnosyl transferase, RhlB, from Pseudomonas aeruginosa (27) (Table 1).…”

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Enhanced antibiotic production by manipulation of the Streptomyces peucetius dnrH and dnmT genes involved in doxorubicin (adriamycin) biosynthesis

Scotti

Hutchinson

1996

J Bacteriol

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Sequence analysis of a 3.4-kb region Streptomyces peucetius daunorubicin (DNR) gene cluster established the presence of the dnrH and dnmT genes. In dnrH mutants, DNR production increased 8.5-fold, compared with that in the wild-type strain, while dnmT mutants accumulated -rhodomycinone (RHO), which normally becomes glycosylated in daunorubicin biosynthesis. Hence, dnmT may be involved in the biosynthesis or attachment of daunosamine to RHO or in the regulation of this process. Since the DnrH protein is similar to known glycosyl transferases, this protein may catalyze the conversion of DNR to its polyglycosylated forms, known as baumycins. Overexpression of dnmT in the wild-type and dnrH mutant strains resulted in a major decrease in RHO accumulation and increase in DNR production.Daunorubicin (DNR) and its C-14 hydroxylated derivative doxorubicin (DXR) are important antitumor anthracycline antibiotics (26) isolated from Streptomyces peucetius ATCC 29050 (2, 5, 7). Studies of DNR and DXR production ( Fig. 1) in this organism have elucidated the organization and regulation of many of the biosynthetic genes (11,12,21,23,29,31), as well as the mechanism of antibiotic resistance (14, 19) and the regulation of DNR and DXR biosynthesis (20,28,34).Here we report the characterization of the dnrH and dnmT genes, which are situated between the dpsH (formerly dnr-ORF7) and dnrE (formerly dnrH) genes in the cluster of DXR biosynthesis genes ( Fig. 2 and 3). Dickens et al. (10) have described homologs of these genes in the Streptomyces sp. strain C5 (dau-ORF2 and -ORF3) but were only able to suggest a possible function for dnrH, as deduced from the DNA sequence. From the results of the expression of the dnrH and dnmT genes in the wild type and a dnrH mutant strain, plus the effects of insertional inactivation of both genes, it seems likely that these two genes govern steps subsequent to the formation of the aglycone ε-rhodomycinone (RHO), a key intermediate of DNR biosynthesis (Fig. 1). Moreover, introduction of dnmT on a high-copy-number plasmid into the dnrH mutant resulted in an 8.5-fold increase in DNR production with a concomitant large decrease in the level of RHO, a useless by product of DNR-producing strains.Sequence analysis of the dnrH and dnmT genes. DNA sequencing of both strands of a 2,162-nucleotide (nt) SphIBamHI fragment by previously reported methods (29) followed by CODONPREFERENCE analysis (9) revealed the C-terminal end of dpsH (the rest of this gene has been sequenced in other work [30]), the complete dnmT gene, and the N-terminal part of dnrH. The remainder of dnrH extends 931 nt into the adjacent 1.3-kb BamHI-AlwNI fragment described by Grimm et al. (13) (Fig. 3). The first ATG codon for dnmT is likely to be located at nt 237 (Fig. 3), even though a probable ribosome binding site is not evident near the 5Ј end of this open reading frame (ORF), and a stop codon (TGA) is located at position 1,752, which suggests that dnmT encodes a 505-amino-acid polypeptide with an M r of 55,433. The dnrH gene has a pr...

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Cloning and characterization of two genes from Streptomyces lividans that confer inducible resistance to lincomycin and macrolide antibiotics

Cited by 84 publications

References 19 publications

Dispensable ribosomal resistance to spiramycin conferred by srmA in the spiramycin producer Streptomyces ambofaciens The EMBL/GenBank accession number for the nucleotide sequence described in this paper is AJ223970.

Dispensable ribosomal resistance to spiramycin conferred by srmA in the spiramycin producer Streptomyces ambofaciens The EMBL/GenBank accession number for the nucleotide sequence described in this paper is AJ223970.

Erythromycin resistance by ribosome modification

Enhanced antibiotic production by manipulation of the Streptomyces peucetius dnrH and dnmT genes involved in doxorubicin (adriamycin) biosynthesis

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