Decay-Initiating Endoribonucleolytic Cleavage by RNase Y Is Kept under Tight Control via Sequence Preference and Subcellular Localisation
Decay-Initiating Endoribonucleolytic Cleavage by RNase Y Is Kept under Tight Control via Sequence Preference and Subcellular Localisation
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DOI：
10.1371/journal.pgen.1005577
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发表时间：
2015-10-01
期刊：
影响因子：
4.5
通讯作者：
Redder, Peter
中科院分区：
文献类型：
--
作者：
Khemici, Vanessa;Prados, Julien;Redder, Peter
细菌依赖于高效的RNA周转，无论是在动态平衡期间，还是在基因表达快速变化时都是如此。然而，关于RNA靶向和衰变中的限速步骤，人们知之甚少。膜锚定核糖核酸酶Y是革兰氏阳性菌的毒力因子。我们已经获得了金黄色葡萄球菌核糖核酸酶Y序列特异性的全景图，使用RNA-seq和新的转录组范围的emote方法。对体内产生的99个内切核糖核酸点进行了精确定位，特别是在RNaseY缺失突变体中半衰期增加最多的七个基因中的六个基因内，以及另外三个编码降解核糖核酸酶的单独转录本中，包括RNaseY本身，这表明存在一个调控网络。我们发现，RNaseY是启动大约100个转录本的主要降解途径所必需的，这些转录物不能被其他核糖核酸酶访问，但由于膜限制和对鸟苷的序列偏好而防止了混杂活性。
Bacteria depend on efficient RNA turnover, both during homeostasis and when rapidly altering gene expression in response to changes. Nevertheless, remarkably few details are known about the rate-limiting steps in targeting and decay of RNA. The membrane-anchored endoribonuclease RNase Y is a virulence factor in Gram-positive pathogens. We have obtained a global picture of Staphylococcus aureus RNase Y sequence specificity using RNA-seq and the novel transcriptome-wide EMOTE method. Ninety-nine endoribonucleolytic sites produced in vivo were precisely mapped, notably inside six out of seven genes whose half-lives increase the most in an RNase Y deletion mutant, and additionally in three separate transcripts encoding degradation ribonucleases, including RNase Y itself, suggesting a regulatory network. We show that RNase Y is required to initiate the major degradation pathway of about a hundred transcripts that are inaccessible to other ribonucleases, but is prevented from promiscuous activity by membrane confinement and sequence preference for guanosines.