Instituto de Biotecnología y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata y CCT-La Plata, CONICET, La Plata 1900, Argentina.
YPF Tecnología S.A. (Y-TEC), Avenida. del Petróleo Argentino s/n (1923), Berisso 1923, Argentina.
Int J Mol Sci. 2024 Feb 10;25(4):2157. doi: 10.3390/ijms25042157.
can live inside soybean root nodules and in free-living conditions. In both states, when oxygen levels decrease, cells adjust their protein pools by gene transcription modulation. PhaR is a transcription factor involved in polyhydroxyalkanoate (PHA) metabolism but also plays a role in the microaerobic network of this bacterium. To deeply uncover the function of PhaR, we applied a multipronged approach, including the expression profile of a mutant at the transcriptional and protein levels under microaerobic conditions, and the identification of direct targets and of proteins associated with PHA granules. Our results confirmed a pleiotropic function of PhaR, affecting several phenotypes, in addition to PHA cycle control. These include growth deficiency, regulation of carbon and nitrogen allocation, and bacterial motility. Interestingly, PhaR may also modulate the microoxic-responsive regulatory network by activating the expression of and repressing , both encoding two transcription factors relevant for microaerobic regulation. At the molecular level, two PhaR-binding motifs were predicted and direct control mediated by PhaR determined by protein-interaction assays revealed seven new direct targets for PhaR. Finally, among the proteins associated with PHA granules, we found PhaR, phasins, and other proteins, confirming a dual function of PhaR in microoxia.
能够生活在大豆根瘤内和自由生活条件下。在这两种状态下,当氧气水平下降时,细胞通过基因转录调节来调整其蛋白质库。PhaR 是一种参与聚羟基烷酸(PHA)代谢的转录因子,但在该细菌的微氧网络中也发挥作用。为了深入揭示 PhaR 的功能,我们采用了一种多管齐下的方法,包括在微氧条件下对突变体进行转录和蛋白质水平的表达谱分析,以及直接靶标和与 PHA 颗粒相关的蛋白质的鉴定。我们的结果证实了 PhaR 的多效性功能,除了控制 PHA 循环外,还影响了几种表型。这些表型包括生长缺陷、碳氮分配的调节以及细菌的运动性。有趣的是,PhaR 还可以通过激活 和 的表达并抑制 的表达来调节微氧响应调控网络, 编码两种与微氧调节相关的转录因子。在分子水平上,预测了两个 PhaR 结合基序,并通过蛋白质相互作用实验证实了 PhaR 的直接控制,揭示了 PhaR 的七个新的直接靶标。最后,在与 PHA 颗粒相关的蛋白质中,我们发现了 PhaR、phasins 和其他蛋白质,证实了 PhaR 在微氧中的双重功能。