Cloning and characterization of an aminoglycoside resistance determinant from Micromonospora zionensis.
The sisomicin-gentamicin resistance methylase (sgm) gene was isolated from Micromonospora zionensis and cloned in Streptomyces lividans. The sgm gene was expressed in Micromonospora melanosporea, where its own promoter was active, and also in Escherichia coli under the control of the lacZ promoter. The complete nucleotide sequence of 1,122 bp and a transcription start point were determined. The sequence contains an open reading frame that encodes a polypeptide of 274 amino acids. The methylation of 30S ribosomal subunits by Sgm methylase accounts adequately for all known resistance characteristics of M. zionensis, but expression of high-level resistance to hygromycin B is background dependent. A comparison of the amino acid sequence of the predicted Sgm protein with the deduced amino acid sequences for the 16S rRNA methylases showed extensive similarity of Grm and significant similarity to KgmB but not to KamB methylase.
Kojic
M
M
Institute of Molecular Genetics and Genetic Engineering, Belgrade, Yugoslavia.
Topisirovic
L
L
Vasiljevic
B
B
D000595:Amino Acid Sequence; D000617:Aminoglycosides / Q000494:pharmacology*; D001426:Bacterial Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D004352:Drug Resistance, Microbial / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D005839:Gentamicins / Q000494:pharmacology; D008780:Methyltransferases / Q000235:genetics*; D008847:Micromonospora / Q000187:drug effects / Q000235:genetics*; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D012853:Sisomicin / Q000494:pharmacology; D013302:Streptomyces / Q000235:genetics
D000617:Aminoglycosides; D001426:Bacterial Proteins; D005839:Gentamicins; D008780:Methyltransferases; C078392:sgm protein, Micromonospora zionensis; C035181:rRNA (adenosine-O-2'-)methyltransferase; D012853:Sisomicin