From a jack-of-all-trades arise masters of few: uncovering the evolutionary patterns and processes driving multigene family functional diversification

专门的代谢物不仅是生活的调味品，它们还介导几乎所有生物体与环境的相互作用。由于它们的重要性，这项拟议的工作旨在了解植物和微生物产生的20万种化学物质是如何进化的。 历史遗传机制的知识，允许现有的代谢产物的生产应允许产生新的代谢产物的蛋白质工程应用于农业，医学，生物工程和制药。为了实现这些研究目标，将对处于不同发展阶段的下一代多元化科学家进行培训并收集数据。至少有100名本科生将从事真实的研究经验，产生新的数据，使他们的研究结果与蛋白质进化和专门的代谢物生物合成可以发表在同行评审的期刊。这一经验有望增加学生进入科学研究事业的可能性。基因组复杂性的起源尚不清楚，但至少部分依赖于基因复制产生多基因家族。该项目将研究基因复制后促进多基因家族功能多样化的因素，这一主题广泛适用于参与生命所需反应的数千种酶。要做到这一点，将研究甲基转移酶的SABATH酶家族，该家族产生植物发育和防御的关键分子。祖先序列复活将被用来直接研究产生现代酶的祖先酶。首先，将进行10，000种酶测定，以通过复活古老的复制前和复制后酶来确定蛋白质功能分歧是否在整个酶家族多样化中是不对称的。其次，两个后代基因在生化功能方面优于一个祖先基因的假设也将得到验证。为此，将测量复制前和复制后酶产生的生化产物的量。第三，祖先的蛋白质晶体结构将产生多个祖先和后代酶在这个家庭和分析比较的情况下，以获得洞察活性位点配置伴随着进化变化的酶底物preference.This奖项反映了NSF的法定使命，并已被认为是值得通过评估使用基金会的智力价值和更广泛的影响审查标准的支持。

Specialized metabolites are not only the spice of life, they mediate nearly all organismal interactions with the environment. Due to their importance, this proposed work aims to understand how the 200,000 chemicals produced by plants and microorganisms evolved. Knowledge of the historical genetic mechanisms allowing for existing metabolites to be produced should allow for the generation of novel metabolites by protein engineering for applications in agriculture, medicine, bioengineering and pharmaceuticals. In order to achieve these research goals, the next generation of diverse scientists at different stages of development will be trained and collect data. At least 100 undergraduate students will be engaged in authentic research experiences that generate novel data so that their findings related to protein evolution and specialized metabolite biosynthesis can be published in peer-reviewed journals. This experience is expected to increase the probability of student recruitment into scientific research careers.The origins of genomic complexity are unclear but at least partly depend on gene duplication to generate multigene families. This project will investigate the factors that promote multigene family functional diversification after gene duplication, a topic of broad applicability to the thousands of enzymes involved in reactions required for life. To do this, the SABATH enzyme family of methyltransferases, which produce critical molecules for plant development and defense, will be studied. Ancestral Sequence Resurrection will be employed to directly study the ancestral enzymes that gave rise to modern-day enzymes. First, 10,000 enzyme assays will be performed to determine whether protein functional divergence is asymmetrical throughout enzyme family diversification by resurrecting ancient pre- and post-duplication enzymes. Second, the hypothesis that two descendant genes are better than one ancestral gene in terms of biochemical function will also be tested. To do this, the amount of biochemical products produced by pre- and post-duplication enzymes will be measured. Third, ancestral protein crystal structures will be generated for multiple ancestral and descendant enzymes in this family and analyzed in a comparative context to gain insight into active site configurations concomitant with evolutionary changes in enzyme substrate preferences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.