Ceizel Borella Germán, Lagares Antonio, Valverde Claudio
Laboratorio de Bioquímica, Microbiología e Interacciones Biológicas en el Suelo, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, Bernal B1876BXD, Buenos Aires, Argentina.
Laboratorio de Bioquímica, Microbiología e Interacciones Biológicas en el Suelo, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, Bernal B1876BXD, Buenos Aires, Argentina
FEMS Microbiol Lett. 2016 May;363(9). doi: 10.1093/femsle/fnw069. Epub 2016 Mar 23.
Small non-coding regulatory RNAs (sRNAs) are key players in post-transcriptional regulation of gene expression. Hundreds of sRNAs have been identified in Sinorhizobium meliloti, but their biological function remains unknown for most of them. In this study, we characterized the expression pattern of the gene encoding the 77-nt sRNA MmgR in S. meliloti strain 2011. A chromosomal transcriptional reporter fusion (PmmgR-gfp) showed that the mmgR promoter is active along different stages of the interaction with alfalfa roots. In pure cultures, PmmgR-gfp activity paralleled the sRNA abundance indicating that mmgR expression is primarily controlled at the level of transcriptional initiation. PmmgR-gfp activity was higher during growth in rhizobial defined medium (RDM) than in TY medium. Furthermore, PmmgR-gfp was induced at 60 min after shifting growing cells from TY to RDM medium, i.e. shorter than the cell doubling time. In defined RDM medium containing NO3 (-), both PmmgR-gfp and MmgR level were repressed by the addition of tryptone or single amino acids, suggesting that mmgR expression depends on the cellular nitrogen (N) status. In silico analysis failed to detect conserved motifs upstream the promoter RNA polymerase binding site, but revealed a strongly conserved motif centered at -28 that may be linked to the observed regulatory pattern by the N source.
小非编码调节RNA(sRNA)是基因表达转录后调控的关键参与者。在苜蓿中华根瘤菌中已鉴定出数百种sRNA,但其中大多数的生物学功能仍不清楚。在本研究中,我们对苜蓿中华根瘤菌2011菌株中编码77个核苷酸的sRNA MmgR的基因的表达模式进行了表征。染色体转录报告融合体(PmmgR-gfp)显示,mmgR启动子在与苜蓿根相互作用的不同阶段均有活性。在纯培养物中,PmmgR-gfp活性与sRNA丰度平行,表明mmgR表达主要在转录起始水平受到控制。在根瘤菌限定培养基(RDM)中生长时,PmmgR-gfp活性高于在TY培养基中的活性。此外,将生长中的细胞从TY培养基转移到RDM培养基后60分钟,PmmgR-gfp被诱导,即短于细胞倍增时间。在含有NO3(-)的限定RDM培养基中,添加胰蛋白胨或单一氨基酸会抑制PmmgR-gfp和MmgR水平,这表明mmgR表达取决于细胞的氮(N)状态。计算机分析未能在启动子RNA聚合酶结合位点上游检测到保守基序,但揭示了一个以-28为中心的高度保守基序,该基序可能与观察到的由氮源引起的调控模式有关。