Fakhr M K, Peñaloza-Vázquez A, Chakrabarty A M, Bender C L
Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma 74078, USA.
J Bacteriol. 1999 Jun;181(11):3478-85. doi: 10.1128/JB.181.11.3478-3485.1999.
Both Pseudomonas aeruginosa and the phytopathogen P. syringae produce the exopolysaccharide alginate. However, the environmental signals that trigger alginate gene expression in P. syringae are different from those in P. aeruginosa with copper being a major signal in P. syringae. In P. aeruginosa, the alternate sigma factor encoded by algT (sigma22) and the response regulator AlgR1 are required for transcription of algD, a gene which encodes a key enzyme in the alginate biosynthetic pathway. In the present study, we cloned and characterized the gene encoding AlgR1 from P. syringae. The deduced amino acid sequence of AlgR1 from P. syringae showed 86% identity to its P. aeruginosa counterpart. Sequence analysis of the region flanking algR1 in P. syringae revealed the presence of argH, algZ, and hemC in an arrangement virtually identical to that reported in P. aeruginosa. An algR1 mutant, P. syringae FF5.32, was defective in alginate production but could be complemented when algR1 was expressed in trans. The algD promoter region in P. syringae (PsalgD) was also characterized and shown to diverge significantly from the algD promoter in P. aeruginosa. Unlike P. aeruginosa, algR1 was not required for the transcription of algD in P. syringae, and PsalgD lacked the consensus sequence recognized by AlgR1. However, both the algD and algR1 upstream regions in P. syringae contained the consensus sequence recognized by sigma22, suggesting that algT is required for transcription of both genes.
铜绿假单胞菌和植物病原菌丁香假单胞菌都能产生胞外多糖藻酸盐。然而,触发丁香假单胞菌中藻酸盐基因表达的环境信号与铜绿假单胞菌不同,铜是丁香假单胞菌中的主要信号。在铜绿假单胞菌中,由algT(sigma22)编码的替代sigma因子和应答调节因子AlgR1是algD转录所必需的,algD是一个编码藻酸盐生物合成途径中关键酶的基因。在本研究中,我们克隆并鉴定了来自丁香假单胞菌的编码AlgR1的基因。丁香假单胞菌AlgR1的推导氨基酸序列与其铜绿假单胞菌对应物具有86%的同一性。对丁香假单胞菌中algR1侧翼区域的序列分析表明,存在argH、algZ和hemC,其排列方式与铜绿假单胞菌中报道的几乎相同。algR1突变体丁香假单胞菌FF5.32在藻酸盐产生方面存在缺陷,但当algR1以反式表达时可以得到互补。丁香假单胞菌中的algD启动子区域(PsalgD)也得到了表征,并显示出与铜绿假单胞菌中的algD启动子有显著差异。与铜绿假单胞菌不同,丁香假单胞菌中algD的转录不需要algR1,并且PsalgD缺乏AlgR1识别的共有序列。然而,丁香假单胞菌中algD和algR1的上游区域都包含sigma22识别的共有序列,这表明algT是这两个基因转录所必需的。