The biosynthetic gene cluster for coronamic acid, an ethylcyclopropyl amino acid, contains genes homologous to amino acid-activating enzymes and thioesterases.
Coronamic acid (CMA), an ethylcyclopropyl amino acid derived from isoleucine, functions as an intermediate in the biosynthesis of coronatine, a chlorosis-inducing phytotoxin produced by Pseudomonas syringae pv. glycinea PG4180. The DNA required for CMA biosynthesis (6.9 kb) was sequenced, revealing three distinct open reading frames (ORFs) which share a common orientation for transcription. The deduced amino acid sequence of a 2.7-kb ORF designated cmaA contained six core sequences and two conserved motifs which are present in a variety of amino acid-activating enzymes, including nonribosomal peptide synthetases. Furthermore, CmaA contained a spatial arrangement of histidine, aspartate, and arginine residues which are conserved in the ferrous active site of some nonheme iron(II) enzymes which catalyze oxidative cyclizations. The deduced amino acid sequence of a 1.2-kb ORF designated cmaT was related to thioesterases of both procaryotic and eucaryotic origins. These data suggest that CMA assembly is similar to the thiotemplate mechanism of nonribosomal peptide synthesis. No significant similarities between a 0.9-kb ORF designated cmaU and other database entries were found. The start sites of two transcripts required for CMA biosynthesis were identified in the present study. pRG960sd, a vector containing a promoterless glucuronidase gene, was used to localize and study the promoter regions upstream of the two transcripts. Data obtained in the present study indicate that CMA biosynthesis is regulated at the transcriptional level by temperature.
Ullrich
M
M
Department of Plant Pathology, Oklahoma State University, Stillwater 74078-9947.
Bender
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CL
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics*; D016254:Mutagenesis, Insertional; D010453:Peptide Synthases / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011549:Pseudomonas / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D013869:Thiolester Hydrolases / Q000235:genetics; D014158:Transcription, Genetic
D000596:Amino Acids; D001426:Bacterial Proteins; C091077:CmaA protein, Pseudomonas syringae; C091074:CmaU protein, Pseudomonas syringae; C043950:1-amino-2-ethylcyclopropane-1-carboxylic acid; D013869:Thiolester Hydrolases; C091073:cmaT protein, Pseudomonas syringae; D010453:Peptide Synthases