Identification of the biosynthetic gene cluster of A-500359s in Streptomyces griseus SANK60196.
A-500359s, produced by Streptomyces griseus SANK60196, are inhibitors of bacterial phospho-N-acetylmuramyl-pentapeptide translocase. They are composed of three distinct moieties: a 5'-carbamoyl uridine, an unsaturated hexuronic acid and an aminocaprolactam. Two contiguous cosmids covering a 65-kb region of DNA and encoding 38 open reading frames (ORFs) putatively involved in the biosynthesis of A-500359s were identified. Reverse transcriptase PCR showed that most of the 38 ORFs are highly expressed during A-500359s production, but mutants that do not produce A-500359s did not express these same ORFs. Furthermore, orf21, encoding a putative aminoglycoside 3'-phosphotransferase, was heterologously expressed in Escherichia coli and Streptomyces albus, yielding strains having selective resistance against A-500359B, suggesting that ORF21 phosphorylates the unsaturated hexuronic acid as a mechanism of self-resistance to A-500359s. In total, the data suggest that the cloned region is involved in the resistance, regulation and biosynthesis of A-500359s.
Funabashi
Masanori
M
Bioengineering Research Group I, Process Technology Research Laboratories, Pharmaceutical Technology Division, Daiichi Sankyo Co. Ltd, Fukushima, Japan.
Nonaka
Koichi
K
Yada
Chieko
C
Hosobuchi
Masahiko
M
Masuda
Nobuhisa
N
Shibata
Tomoyuki
T
Van Lanen
Steven G
SG
D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000494:pharmacology; D001381:Azepines / Q000494:pharmacology; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D015723:Gene Library; D005798:Genes, Bacterial / Q000235:genetics; D005822:Genetic Vectors; D005810:Multigene Family; D016366:Open Reading Frames; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013305:Streptomyces griseus / Q000235:genetics*; D014529:Uridine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000494:pharmacology
C544388:A 500359B; D000900:Anti-Bacterial Agents; D001381:Azepines; D004269:DNA, Bacterial; D014529:Uridine