RNA metabolism and regulation in Streptomyces bacteria

非编码RNA是所有生物体中细胞过程的重要介质。本研究的目的是了解链霉菌sRNA活性的功能和机制。 链霉菌以其代谢多样性和形态复杂性而闻名，产生大量可用于医学（人类和兽医）和农业的代谢产物，包括除草剂，抗癌剂和大多数已知的抗生素。它们也是多细胞生物，可以分化形成不同的组织类型，为在简单的细菌系统中研究细胞分化提供了机会。 ** 利用RNA测序的力量，我们评估了三种进化上不同的链霉菌的sRNA潜力，并确定了25种保守sRNA的核心组。 我们将描述选择sRNA的生物学功能，优先考虑那些影响发育或代谢的sRNA。为了确定sRNA调控靶点，我们将采取多维方法，将体内亲和纯化与转录组学和蛋白质组学策略相结合。将使用体内和体外sRNA-靶标相互作用研究揭示作用机制，并将与旨在探索核糖核酸酶在sRNA介导的调节中的作用及其对RNA代谢的贡献的实验相结合。我们将利用我们对sRNA介导的基因调控的理解，开发旨在工程和操纵链霉菌调控网络的合成策略。** 本研究提案的长期目标是定义新的sRNA调控途径，并将其整合到已知的控制形态分化和代谢产物产生的级联反应中。这项工作将提供链霉菌生物学的基本见解，将揭示细菌基因调控的普遍主题和独特的调控策略，并将与生物技术产业高度相关。

Non-coding RNAs are important mediators of cellular processes in all groups of organisms. The aim of this proposal is to understand the function and mechanism of sRNA activity in the Streptomyces bacteria. The streptomycetes are renowned for their metabolic diversity and morphological complexity, producing a vast array of metabolites with utility in medicine (human and veterinary) and agriculture, including herbicides, anti-cancer agents, and most known antibiotics. They are also multicellular organisms that differentiate to form distinct tissue types, providing an opportunity to study cell differentiation in a simple bacterial system. ******Leveraging the power of RNA-sequencing, we have assessed the sRNA potential of three evolutionarily divergent streptomycetes, and have defined a core group of 25 conserved sRNAs. We will characterize the biological function of select sRNAs, prioritizing those affecting development or metabolism. To identify sRNA regulatory targets, we will take a multi-dimensional approach, integrating in vivo affinity purification with transcriptomic and proteomic strategies. Mechanisms of action will be revealed using in vivo and in vitro sRNA-target interaction studies, and will be coupled with experiments aimed at exploring the role of ribonucleases in sRNA-mediated regulation, and their contributions to RNA metabolism in general. We will exploit our understanding of sRNA-mediated gene regulation to develop synthetic strategies aimed at the engineering and manipulation of Streptomyces regulatory networks. ******The long term objective of this research proposal is to define novel sRNA-regulatory pathways, and integrate these into known cascades governing morphological differentiation and metabolite production. This work will provide fundamental insights into Streptomyces biology, will reveal both universal themes in bacterial gene regulation and unique regulatory strategies, and will be highly relevant to the biotechnology industry.