Nucleotide sequence and transcriptional analysis of the Streptomyces griseus gene (afsA) responsible for A-factor biosynthesis

Abstract

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.