Department of Plant Pathology, China Agricultural University, Beijing, People's Republic of China.
PLoS One. 2012;7(11):e50149. doi: 10.1371/journal.pone.0050149. Epub 2012 Nov 29.
Pseudomonas fluorescens 2P24 is a rhizospheric bacterium that aggressively colonizes the plant roots. It produces the antibiotic 2,4-diacetylphoroglucinol (2,4-DAPG), which contributes to the protection of various crop plants against soil borne diseases caused by bacterial and fungal pathogens. The biosynthesis of 2,4-DAPG is regulated at the transcriptional level in the expression of the phlACBD operon as well as at the posttranscriptional level by the Gac/Rsm signal transduction pathway. However, the detailed mechanism of such regulation is not clear.
METHODOLOGY/PRINCIPAL FINDINGS: In this study, we identified a binding site for the sigma regulator PsrA in the promoter region of the phlA gene. Electrophoretic mobility shift experiments revealed direct and specific binding of PsrA to the phlA promoter region. Consistent with the fact that its binding site locates within the promoter region of phlA, PsrA negatively regulates phlA expression, and its inactivation led to significant increase in 2,4-DAPG production. Interestingly, PsrA also activates the expression of the sigma factor RpoS, which negatively regulates 2,4-DAPG production by inducing the expression of the RNA-binding protein RsmA.
CONCLUSIONS/SIGNIFICANCE: These results suggest that PsrA is an important regulator that modulates 2,4-DAPG biosynthesis at both transcriptional and posttranscriptional levels.
荧光假单胞菌 2P24 是一种根际细菌,它会积极定殖在植物根部。它会产生抗生素 2,4-二乙酰基腐胺(2,4-DAPG),这有助于保护各种作物植物免受由细菌和真菌病原体引起的土传疾病的侵害。2,4-DAPG 的生物合成在 phlACBD 操纵子的转录水平以及 Gac/Rsm 信号转导途径的转录后水平受到调控。然而,这种调控的详细机制尚不清楚。
方法/主要发现:在这项研究中,我们在 phlA 基因的启动子区域鉴定到了 sigma 调控因子 PsrA 的结合位点。电泳迁移率变动实验显示 PsrA 与 phlA 启动子区域的直接和特异性结合。与它的结合位点位于 phlA 启动子区域内的事实一致,PsrA 负调控 phlA 的表达,其失活导致 2,4-DAPG 产量显著增加。有趣的是,PsrA 还激活了 sigma 因子 RpoS 的表达,RpoS 通过诱导 RNA 结合蛋白 RsmA 的表达来负调控 2,4-DAPG 的产生。
结论/意义:这些结果表明,PsrA 是一种重要的调节剂,可在转录和转录后水平上调节 2,4-DAPG 的生物合成。