Nucleotide sequence and transcriptional analysis of the Streptomyces griseus gene (afsA) responsible for A-factor biosynthesis.
The nucleotide sequence of the Streptomyces griseus afsA gene, possibly encoding a key enzyme for A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone) biosynthesis, was determined. The translational initiation codon was identified by introducing out-of-frame mutations at appropriate positions by oligonucleotide-directed mutagenesis. The afsA gene was thus found to code for a protein of 301 amino acid residues and 32.6 kilodaltons whose codon usage pattern was in agreement with the general tendency of Streptomyces genes with an extremely high guanine-plus-cytosine content. High-resolution S1 nuclease mapping indicated that the transcriptional start point was the A residue, the first position of the ATG translational initiation codon.
Horinouchi
S
S
Department of Agricultural Chemistry, University of Tokyo, Japan.
Suzuki
H
H
Nishiyama
M
M
Beppu
T
T
D015107:4-Butyrolactone / Q000031:analogs & derivatives; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D005796:Genes; D005798:Genes, Bacterial; D006133:Growth Substances / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D015183:Restriction Mapping; D013305:Streptomyces griseus / Q000235:genetics*; D014158:Transcription, Genetic
C108224:AfsA protein, Streptomyces griseus; D001426:Bacterial Proteins; D006133:Growth Substances; C025035:A-factor (Streptomyces); D015107:4-Butyrolactone