CAREER: Cracking the Cleavage Code of RNase Y and Its Associated Y-Complex in Firmicutes

这项研究的目的是阐明细菌基因组中决定蛋白质产量的一种神秘密码。这些新信息将有助于解释细菌如何利用遗传信息与它们生活的环境相互作用并改变它们所处的环境，这反过来可能产生从基础生物学到生物技术的广泛影响。参与该项目的研究生将学习开发用于分析大规模数据集的新的高通量测量和统计方法。该项目的教育和宣传部分与研究的跨学科性质相结合，将促进生物学和统计学方面的综合学习体验。假设驱动的研究方法和假设检验统计概念都将以互动的形式教授，促进好奇心驱动的探究和扩大参与。信使RNA是蛋白质合成的模板，它们的寿命决定蛋白质的产量。这项研究将确定导致革兰氏阳性细菌RNA降解的初始切割事件的分子决定因素。新的全基因组技术将被用来揭示不同类别的切割位点，并将通过生物信息学确定它们之间的共性。这项纬度调查将与两项纵向研究相辅相成，使用深度突变扫描和保护的系统发育分析。此外，将进行全基因组筛查，以提供调节核糖核酸酶活性的反式作用因子的第一个全面视图。切割密码及其效应器的定量定义将极大地提高我们对细菌所使用的基因表达程序的理解，并潜在地改变我们为合成生物学设计可预测的遗传电路的能力。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The objective of this research is to elucidate a cryptic code in bacterial genomes that determines the rate of protein production. This new information will help explain how bacteria utilize genetic information to interact with and change the environment they live in, which in turn could have wide ranging impacts, from fundamental biology to biotechnology. Graduate students participating in the project will learn to develop novel high-throughput measurements and statistical methods for analyzing large-scale data sets. Synergistic with the interdisciplinary nature of the research, the education and outreach components of this project will promote an integrated learning experience in biology and statistics. Both hypothesis-driven research methods and hypothesis-testing statistical concepts will be taught in an interactive format that promotes curiosity-driven inquiry and broadens participation.Messenger RNAs are templates for protein synthesis, and their lifetimes determine the yield of proteins. This research will identify the molecular determinants of the initial cleavage event that leads to RNA degradation in Gram-positive bacteria. Novel genome-wide techniques will be leveraged to reveal different classes of cleavage sites, and commonalities among them will be identified through bioinformatics. This latitudinal investigation will be complemented with two longitudinal studies using deep mutational scanning and phylogenetic analysis of conservation. Furthermore, genome-wide screens will be performed to provide the first comprehensive view of trans-acting factors that regulate RNase activities. A quantitative definition of cleavage codes and their effectors will greatly enhance our understanding of gene expression programs employed by bacteria, and potentially transform our ability to design predictable genetic circuits for synthetic biology.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.