Identification of genes encoding for peptide synthetases in the gram-negative bacterium Lysobacter sp. ATCC 53042 and the fungus Cylindrotrichum oligospermum.
Genes encoding for the multifunctional peptide synthetases lysobactin synthetase and peptolide SDZ 214-103 synthetase were identified by hybridization of genomic libraries with oligonucleotides derived from consensus motifs of various genes encoding for delta-(L-alpha amino-adipoyl)-L-cysteinyl-D-valine (ACV) synthetases and gramicidin S synthetase. The sequence of subcloned gene fragments revealed core motifs and a modular structure typical for the family of peptide synthetase genes. A fragment of 4.6 kb of the lysobactin synthetase gene was sequenced and one amino acid activating module was localized. The cloning of lysobactin synthetase was verified by marker-exchange mutagenesis and the lysobactin minus phenotype of the mutant. The sequenced 3.1 kb fragment of peptolide SDZ 214-103 synthetase contained parts of two modules and was highly homologous to corresponding regions of module 6 and 7 of cyclosporin synthetase. Therefore, the localized modules may activate the amino acids threonine and glycine.
Bernhard
F
F
Freie Universität Berlin, Institut für Kristallographie, Germany. fbern@chemie.fuberlin.de
Demel
G
G
Soltani
K
K
Döhren
H V
HV
Blinov
V
V
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D015245:Deoxyribonuclease BamHI / Q000378:metabolism; D005658:Fungi / Q000201:enzymology / Q000235:genetics*; D005798:Genes, Bacterial; D005800:Genes, Fungal; D006090:Gram-Negative Bacteria / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D015345:Oligonucleotide Probes; D010453:Peptide Synthases / Q000235:genetics*
D004274:DNA, Recombinant; D015345:Oligonucleotide Probes; D015245:Deoxyribonuclease BamHI; D010453:Peptide Synthases