Centro Nacional de Biotecnología, CSIC, Darwin 3, 28049, Madrid, Spain.
Sci Rep. 2017 Nov 22;7(1):16016. doi: 10.1038/s41598-017-16091-0.
Gene expression is determined by critical processes such as RNA synthesis and degradation. Ribonucleases participate in the coordinated and differential decay of messenger RNAs. We describe a suitable method of normalization and calculation of mRNAs half-life values quantified by RNA-Seq. We determined the mRNA half-lives of more than 2000 genes in Stenotrophomonas maltophilia D457 and in an isogenic RNase G deficient mutant. Median half-lives were 2,74 and 3 min in the wild-type and the rng-deficient strain, respectively. The absence of RNase G resulted in an overall enhancement of mRNA half-life times, showing that many RNAs are targets of RNase G in S. maltophilia. Around 40 genes are likely to be regulated directly by RNase G since their half-lives were more than two-fold higher in the rng-deficient mutant. Gene length, GC content or expression levels did not correlate with mRNAs lifetimes, although groups of genes with different functions showed different RNA half-lives. Further, we predicted 1542 gene pairs to be part of the same operons in S. maltophilia. In contrast to what was described for other bacteria, our data indicate that RNase G has a global role in mRNA stability and consequently in the regulation of S. maltophilia gene expression.
基因表达是由 RNA 合成和降解等关键过程决定的。核糖核酸酶参与信使 RNA 的协调和差异降解。我们描述了一种适用于通过 RNA-Seq 定量的 mRNA 半衰期值的归一化和计算方法。我们确定了嗜麦芽寡养单胞菌 D457 及其同源 RNase G 缺陷突变体中超过 2000 个基因的 mRNA 半衰期。野生型和 rng 缺陷型菌株的 mRNA 半衰期中位数分别为 2.74 和 3 分钟。缺乏 RNase G 导致 mRNA 半衰期总体延长,表明许多 RNA 是嗜麦芽寡养单胞菌中 RNase G 的靶标。大约 40 个基因可能受到 RNase G 的直接调控,因为它们在 rng 缺陷型突变体中的半衰期是野生型的两倍以上。基因长度、GC 含量或表达水平与 mRNAs 寿命没有相关性,尽管具有不同功能的基因群显示出不同的 RNA 半衰期。此外,我们预测了嗜麦芽寡养单胞菌中有 1542 对基因对是同一操纵子的一部分。与其他细菌的描述相反,我们的数据表明,RNase G 在 mRNA 稳定性和随后的嗜麦芽寡养单胞菌基因表达调控中具有全局作用。