Expression of the phosphonoalanine-degradative gene cluster from Variovorax sp Pal2 is induced by growth on phosphonoalanine and phosphonopyruvate
Expression of the phosphonoalanine-degradative gene cluster from Variovorax sp Pal2 is induced by growth on phosphonoalanine and phosphonopyruvate
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DOI：
10.1111/j.1574-6968.2008.01477.x
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发表时间：
2009-03-01
影响因子：
2.1
通讯作者：
Quinn, John P.
中科院分区：
文献类型：
--
作者：
Kulakova, Anna N.;Kulakov, Leonid A.;Quinn, John P.
在Variovorax sp.，Pal2中发现的磷酸丙酮酸水解酶(Pala)是一种新的碳磷键裂解酶，即使在高水平的磷存在下也能表达，从而使磷酸丙酮酸成为唯一的碳源和能源。对Pala基因邻近区域的分析表明，存在构成2-氨基-3-膦丙酸(膦丙氨酸)降解操纵子的五个结构基因。逆转录-聚合酶链式反应(RT-PCR)实验表明，操纵子中的5个基因均被转录为一个单一的mRNA，并且它们的转录受磷酸丙氨酸或磷酸丙酮酸的诱导。构建了磷酸丙氨酸降解操纵子的调节区与GFP基因的转录融合。表达分析表明，LysR型调节子(由palR基因编码)的存在对于操纵子结构基因的转录是必不可少的。在来自Alpha、Beta和GammaProteb细菌的6种细菌的基因组测序中发现了相似的基因簇，元基因组文库分析表明，与Pala相关的序列在海洋环境中广泛分布。
The phosphonopyruvate hydrolase (PalA) found in Variovorax sp., Pal2, is a novel carbon-phosphorus bond cleavage enzyme, which is expressed even in the presence of high levels of phosphate, thus permitting phosphonopyruvate to be used as the sole carbon and energy source. Analysis of the regions adjacent to the palA gene revealed the presence of the five structural genes that constitute the 2-amino-3-phosphonopropionic acid (phosphonoalanine)-degradative operon. Reverse transcriptase-PCR (RT-PCR) experiments demonstrated that all five genes in the operon are transcribed as a single mRNA and that their transcription is induced by phosphonoalanine or phosphonopyruvate. Transcriptional fusions of the regulatory region of the phosphonoalanine degradative operon with the gfp gene were constructed. Expression analysis indicated that the presence of a LysR-type regulator (encoded by the palR gene) is essential for the transcription of the structural genes of the operon. Similar gene clusters were found in the sequenced genomes of six bacterial species from the Alpha-, Beta- and Gammaproteobacteria, and analysis of metagenomic libraries revealed that sequences related to palA are widely spread in the marine environment.