Accessing the hidden biosynthetic capabilities of fungi

Parent Grant GM112739项目摘要 丝状真菌产生大量的次生代谢物(SM)，其生物活性是 这对医学和农业的进步至关重要。例如，真菌SM表现出细胞抑制作用， 免疫抑制、降脂或抗菌性能。基因组测序的快速进展 丝状真菌发现了大量未知的假定生物合成途径 代谢物，表明发现新化合物和活性的巨大潜力。然而， 对真菌BGC多样性的充分表征存在重大障碍：(A)合成了许多SM 由生物信息学算法无法识别的生物合成基因簇(BGC)，(B)许多 BGC在标准的实验室条件下不表达(例如，隐蔽的BGC)，以及(C)某些基因 生物合成途径并不都是聚集的，涉及一个以上的基因座。此外，(D)几乎没有 对功能性BGC发生机制的理解。我们最近的结果清楚地表明，非正则的 BGC揭示了真正的新结构或不寻常的生物化学，即特定的真菌分化信号 诱导全球BGC表达，重组热点产生BGC多样性。在这笔赠款中， 我们将(I)描述异氰化物合成酶(ICS)BGC，一个最近发现的非正则家族 真菌BGC不能被当前的软件算法识别，我们已经有了初步结果 展示新的和令人兴奋的生物化学，(Ii)使用真菌分化信号进行转录 在不同的真菌分类群中识别“看不见的”BGC和(Iii)解决这样的假设 重组和水平转移的基因组“热点”催生了新的BGC。基于 假设非正则基因簇特别可能产生具有高 结构和功能的新颖性程度，以及能够识别活跃的BGC的两个进步以及如何 BGC是形成的，我们的工具集比较转录学，高级内源和异源 表达平台和最近开发的比较代谢组学平台将提供 通过扩展生物学、遗传学的新结构、生物合成途径和生物活性 和真菌BGC的化学。此外，对BGC成因的洞察和全球BGC诱导的识别 信号提供了真正新的过程，以进一步扩大真菌BGC的发现和功能范围 注释。

PROJECT SUMMARY FOR PARENT GRANT GM112739 Filamentous fungi produce a vast universe of secondary metabolites (SM) with biological activities that are of central importance for progress in medicine and agriculture. For example, fungal SMs exhibit cytostatic, immunosuppressant, lipid lowering, or antimicrobial properties. Rapid progress in sequencing the genomes of filamentous fungi has revealed a very large number of putative biosynthetic pathways with no known metabolites, suggesting a vast potential for the discovery of new compounds and activities. However, significant impediments to full characterization of fungal BGC diversity exist: (a) many SMs are synthesized by ‘non-canonical’ biosynthetic gene clusters (BGCs) not recognized by bioinformatic algorithms, (b) many BGCs are not expressed in standard laboratory conditions (e.g. ‘cryptic’ BGCs), and (c) genes for some biosynthetic pathways are not all clustered and involve more than one locus. Further, there is (d) little understanding of the genesis of functional BGCs. Our recent results clearly indicate that non-canonical BGCs reveal genuinely novel structures or unusual biochemistry, that specific fungal differentiation signals induce global BGC expression and that “hot spots” of recombination generate BGC diversity. In this grant, we will (i) characterize isocyanide synthase (ICS) BGCs, a recently discovered family of noncanonical fungal BGCs not recognized by current software algorithms for which we have preliminary results demonstrating new and exciting biochemistry, (ii) use a fungal differentiation signal for transcriptomic identification of ‘invisible' BGCs across diverse fungal taxa and (iii) address the hypothesis that genomic “hot spots” of recombination and horizontal transfer give birth to new BGCs. Based on the premise that non-canonical gene clusters are particularly likely to produce chemical entities with a high degree of structural and functional novelty coupled with two advances able to identify active BGCs and how BGC are formed, our tool set of comparative transcriptomics, advanced endogenous and heterologous expression platforms and a recently developed platform for comparative metabolomics will provide a wealth of new structures, biosynthetic pathways, and biological activities through expansion of the biology, genetics and chemistry of fungal BGCs. Moreover, insight into BGC genesis and identification of global BGC induction signals present genuinely new processes to further the scope of fungal BGC discovery and functional annotation.