Characterization of virginiamycin S biosynthetic genes from Streptomyces virginiae.
Streptomyces virginiae produces -butyrolactone autoregulators (virginiae butanolide, VB), which control the biosynthesis of virginiamycin M1 and S. A 6.3-kb region downstream of the virginiamycin S (VS)-resistance operon in S. virginiae was sequenced, and four plausible open reading frames (ORFs) (visA, 1,260 bp; visB, 1,656 bp; visC, 888 bp; visD, 1209 bp) were identified. Homology analysis revealed significant similarities with enzymes involved in the biosynthesis of cyclopeptolide antibiotics: VisA (53% identity, 65% similarity) to -lysine 2-aminotransferase (NikC) of nikkomycin D biosynthesis, VisB (66% identity, 72% similarity) to 3-hydroxypicolinic acid:AMP ligase of pristinamycin I biosynthesis, VisC (48% identity, 59% similarity) to lysine cyclodeaminase of ascomycin biosynthesis, and VisD (43% identity, 56% similarity) to erythromycin C-22 hydroxylase of erythromycin biosynthesis. Northern blotting as well as high-resolution S1 analysis of the ORFs revealed that they were transcribed as two bicistronic transcripts, namely 3.0-kb visB-visA and another 2.7-kb visC-visD transcript, with promoters locating upstream of visB and visC, respectively. Transcription of the two operons was observed only 1 h after the VB production, which was 2 h before the virginiamycin production. Furthermore, prompt induction of the transcription was observed as a result of external VB addition, suggesting that the expression of the two operons was under the control of VB.
Namwat
Wises
W
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Kamioka
Yuji
Y
Kinoshita
Hiroshi
H
Yamada
Yasuhiro
Y
Nihira
Takuya
T
D000642:Ammonia-Lyases / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D023061:Gene Order; D005798:Genes, Bacterial / Q000235:genetics*; D008025:Ligases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D000637:Transaminases / Q000235:genetics / Q000378:metabolism; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D014769:Virginiamycin / Q000031:analogs & derivatives / Q000096:biosynthesis* / Q000378:metabolism
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C456604:VisA protein, Streptomyces virginiae; C456605:VisB protein, Streptomyces virginiae; C456606:VisC protein, Streptomyces virginiae; C456607:VisD protein, Streptomyces virginiae; D014769:Virginiamycin; D003577:Cytochrome P-450 Enzyme System; D000637:Transaminases; C113111:lysine 2-aminotransferase; D000642:Ammonia-Lyases; C109507:lysine cyclodeaminase; D008025:Ligases