Grupo de Ecología Genética de la Rizosfera, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas (CSIC), Granada, Spain.
PLoS One. 2013 Jul 15;8(7):e68147. doi: 10.1371/journal.pone.0068147. Print 2013.
The legume symbiont Sinorhizobium meliloti expresses a plethora of small noncoding RNAs (sRNAs) whose function is mostly unknown. Here, we have functionally characterized two tandemly encoded S. meliloti Rm1021 sRNAs that are similar in sequence and structure. Homologous sRNAs (designated AbcR1 and AbcR2) have been shown to regulate several ABC transporters in the related α-proteobacteria Agrobacterium tumefaciens and Brucella abortus. In Rm1021, AbcR1 and AbcR2 exhibit divergent unlinked regulation and are stabilized by the RNA chaperone Hfq. AbcR1 is transcribed in actively dividing bacteria, either in culture, rhizosphere or within the invasion zone of mature alfalfa nodules. Conversely, AbcR2 expression is induced upon entry into stationary phase and under abiotic stress. Only deletion of AbcR1 resulted into a discrete growth delay in rich medium, but both are dispensable for symbiosis. Periplasmic proteome profiling revealed down-regulation of the branched-chain amino acid binding protein LivK by AbcR1, but not by AbcR2. A double-plasmid reporter assay confirmed the predicted specific targeting of the 5'-untranslated region of the livK mRNA by AbcR1 in vivo. Our findings provide evidences of independent regulatory functions of these sRNAs, probably to fine-tune nutrient uptake in free-living and undifferentiated symbiotic rhizobia.
豆科植物共生菌根瘤农杆菌表达大量的小非编码 RNA(sRNA),但其功能大多未知。在这里,我们对串联编码的 S. meliloti Rm1021 sRNA 进行了功能表征,这些 sRNA 在序列和结构上相似。同源 sRNA(命名为 AbcR1 和 AbcR2)已被证明调节相关α变形菌根瘤农杆菌和布鲁氏菌中的几种 ABC 转运蛋白。在 Rm1021 中,AbcR1 和 AbcR2 表现出不同的非连锁调节,并由 RNA 伴侣 Hfq 稳定。AbcR1 在活跃分裂的细菌中转录,无论是在培养物、根际还是在成熟紫花苜蓿根瘤的入侵区。相反,AbcR2 的表达在进入静止期和非生物胁迫下诱导。只有删除 AbcR1 才会导致丰富培养基中的生长明显延迟,但两者对于共生都是可有可无的。周质蛋白组分析显示 AbcR1 下调支链氨基酸结合蛋白 LivK,但 AbcR2 没有。双质粒报告基因测定证实 AbcR1 在体内对 livK mRNA 的 5'-非翻译区进行了预测的特异性靶向。我们的研究结果提供了这些 sRNA 独立调控功能的证据,可能是为了精细调节游离和未分化共生根瘤菌的营养吸收。