Molecular and functional validation of PA0982.1 regulation of the rhl QS system in P. aeruginosa. (IMAGE)
Caption
(A) Predicted RNA‒RNA interaction site between PA0982.1 and the rhlI 5' UTR. (B) RNA EMSA of PA0982.1 binding to the rhlI 5' UTR in vitro, with increasing PA0982.1 concentrations (0, 2, 4, 8 μM) shifting the complex formation, alongside free rhlI 5' UTR and PA0982.1 sRNA. (C) Protein‒RNA EMSA interaction between PA0982.1 and Hfq, with increasing PA0982.1 concentrations forming a PA0982.1-Hfq complex, alongside free PA0982.1 RNA. (D) Quantification of C4-HSL production levels in the sRNA PA0982.1 mutant (ΔPA0982.1) and complemented strains (ΔPA0982.1+), measured relative to wild-type (WT) PAO1. (E) Relative mRNA expression levels of rhlR in WT, ΔPA0982.1, and complemented strains, determined by RT‒qPCR. (F) PYO production in WT, ΔPA0982.1, and complemented strains measured at OD695. (G) Visual representation of PYO production in culture media of WT, ΔPA0982.1, and ΔPA0982.1+. (H) Relative mRNA expression levels of phzF1 in WT, ΔPA0982.1, and complemented strains, determined by RNA-seq and validated by RT‒qPCR. (I) Working mechanism of sRNA PA0982.1 in regulating the rhl QS system and pyocyanin production.
Credit
Tianmin Li, Beifang Lu, Fang Chen, Letong Xu, Yizhou Zhang, Fangzhou Xie, Yue Sun, Chunyan Yao, Jiadai Huang, Tianlong He, Youyue Li, Yiqing Ding, Haiyan Hu, Shaojun Tang, Xin Deng
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