title: Characterization of the syringomycin synthetase gene cluster. A link between prokaryotic and eukaryotic peptide synthetases.
abstract: With this work we have completed the characterization of the syringomycin synthetase gene cluster. In particular, by sequencing additional 28.5 kilobase pairs we show that the nine modules involved in the binding of the nine amino acids of syringomycin are localized on SyrB and SyrE, with SyrE carrying eight modules. The recombinant SyrB and the first and second modules of SyrE (SyrE1 and SyrE2) have been expressed in Escherichia coli and purified. The biochemical data indicate that SyrB binds threonine, the putative precursor of the last amino acid of syringomycin, whereas SyrE1 and SyrE2 bind serine, the first and the second amino acids of syringomycin, respectively. On the basis of the sequence analysis and the biochemical data presented here, it appears that syringomycin synthetase is unique among peptide synthetases in that its genetic organization does not respect the "colinearity rule" according to which the order of the amino acid binding modules along the chromosome parallels the order of the amino acids on the peptide. This feature, together with the absence of a single transcription unit and the absence of epimerase-like domains make syringomycin synthetase more related to the eukaryotic peptide synthetases than to the bacterial counterparts.
authors.affiliation: Department of Molecular Biology, Chiron S.p.A., Via Fiorentina, 1 53100 Siena, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Guenzi
authors.forename: G
authors.initials: G
authors.lastname: Galli
authors.forename: I
authors.initials: I
authors.lastname: Grgurina
authors.forename: D C
authors.initials: DC
authors.lastname: Gross
authors.forename: G
authors.initials: G
authors.lastname: Grandi
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D017931:DNA Primers; D005057:Eukaryotic Cells / Q000201:enzymology; D008969:Molecular Sequence Data; D005810:Multigene Family; D010453:Peptide Synthases; D011387:Prokaryotic Cells / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D017931:DNA Primers; D010453:Peptide Synthases; C094582:SyrC protein, Pseudomonas syringae