Department of Chemistry, Biochemistry and Pharmaceutical Sciences, University of Bern, Freiestrasse 3, 3012 Bern, Switzerland.
Graduate School for Cellular and Biomedical Sciences, University of Bern, 3012 Bern, Switzerland.
Int J Mol Sci. 2021 Feb 8;22(4):1703. doi: 10.3390/ijms22041703.
Almost two-thirds of the microbiome's biomass has been predicted to be in a non-proliferating, and thus dormant, growth state. It is assumed that dormancy goes hand in hand with global downregulation of gene expression. However, it remains largely unknown how bacteria manage to establish this resting phenotype at the molecular level. Recently small non-protein-coding RNAs (sRNAs or ncRNAs) have been suggested to be involved in establishing the non-proliferating state in bacteria. Here, we have deep sequenced the small transcriptome of in the exponential and stationary phases and analyzed the resulting reads by a novel biocomputational pipeline STARPA (Stable RNA Processing Product Analyzer). Our analysis reveals over 12,000 small transcripts enriched during both growth stages. Differential expression analysis reveals distinct sRNAs enriched in the stationary phase that originate from various genomic regions, including transfer RNA (tRNA) fragments. Furthermore, expression profiling by Northern blot and RT-qPCR analyses confirms the growth phase-dependent expression of several enriched sRNAs. Our study adds to the existing repertoire of bacterial sRNAs and suggests a role for some of these small molecules in establishing and maintaining stationary phase as well as the bacterial stress response. Functional characterization of these detected sRNAs has the potential of unraveling novel regulatory networks central for stationary phase biology.
微生物组的生物量中,预计有近三分之二处于非增殖状态,也就是休眠状态。人们认为休眠与基因表达的全面下调是相伴而生的。然而,细菌如何在分子水平上设法建立这种休眠表型在很大程度上仍然未知。最近,人们提出,小非蛋白编码 RNA(sRNA 或 ncRNA)可能参与了细菌非增殖状态的建立。在这里,我们对处于指数生长期和静止期的 进行了小转录组深度测序,并通过一种新的生物计算管道 STARPA(稳定 RNA 处理产物分析器)对得到的读数进行了分析。我们的分析揭示了在两个生长阶段都富集的超过 12000 个小转录本。差异表达分析显示,在静止期富集的特定 sRNA 来源于各种基因组区域,包括转移 RNA (tRNA) 片段。此外,通过 Northern blot 和 RT-qPCR 分析的表达谱证实了几个富集 sRNA 的生长阶段依赖性表达。我们的研究增加了细菌 sRNA 的现有目录,并表明其中一些小分子在建立和维持静止期以及细菌应激反应中发挥作用。对这些检测到的 sRNA 的功能表征有可能揭示出对静止期生物学至关重要的新型调控网络。