Impact of pnpR, a LysR-type regulator-encoding gene, on the cellular processes of Pseudomonas putida DLL-E4
Impact of pnpR, a LysR-type regulator-encoding gene, on the cellular processes of Pseudomonas putida DLL-E4
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LysR 型调节编码基因 pnpR 对恶臭假单胞菌 DLL-E4 细胞过程的影响
DOI：
10.1093/femsle/fnw110
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发表时间：
2016-06-01
影响因子：
2.1
通讯作者：
Cui, Zhongli
中科院分区：
文献类型：
--
作者：
Chen, Qiongzhen;Tu, Hui;Cui, Zhongli
LysR型转录调节因子（LTTR）调节细菌中的各种细胞过程。pnpR是一个参与调节对苯二酚（HQ）降解的LTTR编码基因，本文从生理、生化和分子水平研究了其对恶臭假单胞菌DLL-E4细胞过程的影响。逆转录聚合酶链反应显示，pnpR正调控其自身的表达和pnpC 1C 2DECX 1X 2操纵子的表达;此外，pnpR部分调节pnpA的表达时，恶臭假单胞菌生长对硝基苯酚（PNP）或HQ。菌株DLL-E4和DLL-Delta pnpR表现出相似的细胞形态和生长速率。转录组分析表明，pnpR调节的基因的表达，除了那些参与PNP降解。相对于菌株DLL-E4，在菌株DLL-Delta pnpR中共有20个基因上调，19个基因下调至少2倍。生物信息学分析显示，假定的PnpR结合位点位于参与PNP降解，碳分解代谢产物抑制和其他细胞过程的基因的上游区域。生物活性测定结果表明，DLL-Delta pnpR菌株对L-天冬氨酸、L-组氨酸、L-焦谷氨酸、L-丝氨酸、γ-氨基丁酸、D，L-乳酸、D-糖二酸、琥珀酸和L-丙氨酰胺的利用率提高了1.3倍以上，表明pnpR在碳源利用中起着潜在的负调控作用。
LysR-type transcriptional regulators (LTTRs) regulate various cellular processes in bacteria. pnpR is an LTTR-encoding gene involved in the regulation of hydroquinone (HQ) degradation, and its effects on the cellular processes of Pseudomonas putida DLL-E4 were investigated at the physiological, biochemical and molecular levels. Reverse transcription polymerase chain reaction revealed that pnpR positively regulated its own expression and that of the pnpC1C2DECX1X2 operon; additionally, pnpR partially regulated the expression of pnpA when P. putida was grown on para-nitrophenol (PNP) or HQ. Strains DLL-E4 and DLL-Delta pnpR exhibited similar cellular morphologies and growth rates. Transcriptome analysis revealed that pnpR regulated the expression of genes in addition to those involved in PNP degradation. A total of 20 genes were upregulated and 19 genes were downregulated by at least 2-fold in strain DLL-Delta pnpR relative to strain DLL-E4. Bioinformatic analysis revealed putative PnpR-binding sites located in the upstream regions of genes involved in PNP degradation, carbon catabolite repression and other cellular processes. The utilization of L-aspartic acid, L-histidine, L-pyroglutamic acid, L-serine, gamma-aminobutyric acid, D,L-lactic acid, D-saccharic acid, succinic acid and L-alaninamide was increased at least 1.3-fold in strain DLL-Delta pnpR as shown by BIOLOG assays, indicating that pnpR plays a potential negative regulation role in the utilization of carbon sources.