Experiment based genome mining of ribosomal natural products

描述（由申请人提供）：核糖体编码的天然产物曾被认为具有有限的结构多样性，并且在微生物中不常见。然而，在过去的几年里，这种观点已经改变，由于发现率增加的RNP拥有新描述的结构基序以前归因于他们的非核糖体对应物。几乎每一个测序的基因组，包括无脊椎动物，都含有生物合成核糖体编码的、后修饰的天然产物的遗传能力，所述天然产物例如羊毛硫抗生素、细菌素、微菌素、蓝藻菌素、硫肽和套索肽，从而使得这类未被重视的天然产物可能是所有自然界中最主要的。然而，缺乏的是一种系统的方法来收获这类无处不在的天然产物，并评估其独特的生物合成能力。与以系统方式表征RNP相关的困难可以归因于它们不仅落在大多数治疗筛选程序的范围之外，而且由于它们的较大尺寸、结构多样性和异常数量的翻译后修饰，也落在代谢组学或蛋白质组学方法的范围之外。 该提案概述了开发战略，以创建一套工具，通过基于质谱的基因组挖掘利用核糖体编码的天然产物的生物合成潜力。作为所提出的工作的结果而创建的技术和方法不仅对于检测治疗性先导化合物是重要的，而且对于有效表征由病原性细菌（例如金黄色葡萄球菌、蜡状芽孢杆菌和艰难梭菌）分泌的核糖体编码的毒素以及由高等真核生物（例如海洋蜗牛、灵长类动物和人类）产生的防御素也是重要的。 公共卫生相关性：这项工作概述了一种方法，实验地雷基因组序列的遗传编码分子控制生物学。这些分子是治疗开发的重要先导化合物。

DESCRIPTION (provided by applicant): Ribosomally encoded natural products were once thought to be of limited structural diversity and uncommon amongst microbes. Over the past few years, however, this viewpoint has changed due to the increased discovery rate of RNPs possessing newly described structural motifs previously ascribed to their nonribosomal counterparts. Nearly every sequenced genome, including invertebrates, contains the genetic capacity to biosynthesize ribosomally- encoded, post-translationally modified natural products such as lantibiotics, bacteriocins, microcins, cyanobactins, thiopeptides, and lasso peptides, thereby making this class of underappreciated natural products perhaps the most dominant in all of nature. What is lacking, however, is a systematic approach to harvest this ubiquitous class of natural products and assess their unique biosynthetic capacity. The difficulty associated with characterizing RNPs in a systematic fashion can be attributed to their falling outside the scope of not only most therapeutic screening programs but also metabolomic or proteomic approaches due to their larger size, structural diversity and extraordinary number of post-translational modifications. This proposal outlines the developmental strategies to create a set of tools for harnessing the biosynthetic potential of ribosomally encoded natural products through mass spectrometry based genome mining. The techniques and methodologies created as a result of the proposed work will not only be important for the detection of therapeutic lead compounds, but also for the efficient characterization of ribosomally encoded toxins secreted by pathogenic bacteria such as Staphylococcus aureus, Bacillus cereus and Clostridium difficile as well as defensins produced by higher eukaryotes such as marine snails, primates and humans. PUBLIC HEALTH RELEVANCE: This work outlines an approach that experimentally mines genome sequences for genetically encoded molecules that control biology. Such molecules serve as great lead compounds for therapeutic development.