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首页> 美国卫生研究院文献>Journal of Bacteriology >Identification of the syr-syp Box in the Promoter Regions of Genes Dedicated to Syringomycin and Syringopeptin Production by Pseudomonas syringae pv. syringae B301D
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Identification of the syr-syp Box in the Promoter Regions of Genes Dedicated to Syringomycin and Syringopeptin Production by Pseudomonas syringae pv. syringae B301D

机译：丁香假单胞菌pv专用于丁香霉素和丁香肽素的基因的启动子区的syr-syp盒的鉴定。丁香科B301D

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The phytotoxins syringopeptin and syringomycin are synthesized by nonribosomal peptide synthetases which are encoded by the syringomycin (syr) and syringopeptin (syp) genomic island of Pseudomonas syringae pv. syringae. Previous studies demonstrated that expression of the syr-syp genes was controlled by the salA-syrF regulatory pathway, which in turn was induced by plant signal molecules. In this study, the 132-kb syr-syp genomic island was found to be organized into five polycistronic operons along with eight individual genes based on reverse transcriptional PCR and bioinformatic analysis. The transcriptional start sites of the salA gene and operons III and IV were located 63, 75, and 104 bp upstream of the start codons of salA, syrP, and syrB1, respectively, using primer extension analysis. The predicted −10/−35 promoter region of operon IV was confirmed based on deletion and site-directed mutagenesis analyses of the syrB1::uidA reporter with β-glucuronidase assays. A 20-bp conserved sequence (TGtCccgN6cggGaCA, termed the syr-syp box) with dyad symmetry around the −35 region was identified via computer analysis for the syr-syp genes/operons responsible for biosynthesis and secretion of syringomycin and syringopeptin. Expression of the syrB1::uidA fusion was decreased 59% when 6 bp was deleted from the 5′ end of the syr-syp box in the promoter region of operon IV. These results demonstrate that the conserved promoter sequences of the syr-syp genes contribute to the coregulation of syringomycin and syringopeptin production.
机译：植物毒素syringopeptin和syringomycin由非核糖体肽合成酶合成，该合成酶由丁香假单胞菌pv的syringomycin（syr）和syringopeptin（syp）基因组岛编码。丁香科。先前的研究表明，syr-syp基因的表达受salA-syrF调控途径控制，而后者又由植物信号分子诱导。在这项研究中，基于逆转录PCR和生物信息学分析，发现132kb的syr-syp基因岛被组织为五个多顺反子操纵子以及八个个体基因。使用引物延伸分析，salA基因和操纵子III和IV的转录起始位点分别位于salA，syrP和syrB1起始密码子上游63、75和104 bp。基于syrB1 :: uidA报告基因的缺失和定点诱变分析，并用β-葡糖醛酸糖苷酶测定法确认了操纵子IV的预期-10 / -35启动子区域。通过计算机分析负责生物合成和分泌丁香霉素和丁香肽素的syr-syp基因/操纵子，鉴定了在-35区域周围具有dyad对称性的20 bp保守序列（TGtCccgN6cggGaCA，称为syr-syp盒）。当从操纵子IV的启动子区域中的 syr-syp 框的5'末端删除6 bp时，syrB1 :: uidA 融合体的表达降低59％。这些结果表明 syr-syp 基因的保守启动子序列有助于syringomycin和syringopeptin生产的调控。

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