Optogenetic interrogation of B. subtilis stress-response network dynamics

细菌必须在饥饿和抗生素暴露等环境压力下生存。这些引发压力的条件导致细菌行为和生理学的深刻变化。其中一些变化具有经济和健康影响，因为应激反应与抗应激生物膜的形成、毒性、抗生素耐受性/抗性和持久性有关。该项目的重点是控制细菌应激反应的调控途径。这些结果将为细菌如何在压力下生存提供新的线索，并可能导致合成生物学的新技术进步和急需的抗菌药物。将为全国各地的高中生开发一个新的教育模块。本教育模块将向学生介绍细菌的应激反应以及它们在社会中的重要性。基因调控网络（GRNs）控制着细胞命运决策的基本生物过程。现在很清楚，GRNs利用动态转录因子活性模式作为基本的调控策略。然而，GRNs解码动态转录因子信号的机制通常还不清楚。本项目将使用一种新的光遗传学方法来揭示两种模式B。枯草杆菌应激反应网络解码其主要转录因子调节子的时变活性模式。它将揭示这些途径发挥作用的新机制，并对细菌如何在压力下生存产生新的见解。这些结果还应该提供新的遗传设计原则，这些原则可能是高等生物细胞命运决定的基础。由于动态GRN信号对人类是保守的，这项工作将促进对生命运作规则的新理解。该项目将创建一个新的“Sporograph”教育模块，教导学生细胞如何做出决定。实验模块将以实验室B的廉价版本为基础。本实验室开发的枯草芽孢杆菌光遗传学系统，允许学生通过施加恒定或振荡光输入在细菌琼脂平板上诱导可见的孢子形成和生物膜报告蛋白。在此项目期间，Sporograph模块将惠及数千名学生，并将在未来的努力中进一步扩大。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Bacteria must survive environmental stresses such as starvation and antibiotic exposure. These stress-invoking conditions lead to profound changes in bacterial behavior and physiology. Some of these changes have economic and health implications, as stress responses have been linked to the formation of stress-resistant biofilms, virulence, antibiotic tolerance/resistance and persistence. This project focuses on the regulatory pathways that control stress responses in bacteria. The results will shed new light into how bacteria survive stresses, and could lead to new technological advances in synthetic biology and to sorely-needed antimicrobial drugs. A new educational module will be developed for high-school students across the country. This educational module will teach students about bacterial stress responses and why they are important across society.Gene regulatory networks (GRNs) control the fundamental biological process of cell-fate decision-making. It is now clear that GRNs utilize dynamical transcription factor activity patterns as a fundamental regulatory strategy. However, the mechanisms by which GRNs decode dynamical transcription factor signals are generally not understood. This project will use a new optogenetic method to reveal how two model B. subtilis stress-response networks decode time-varying activity patterns of their major transcription factor regulators. It will reveal new mechanisms by which these pathways function and generate new insights into how bacteria survive stresses. The results should also provide new genetic design principles that may underlie cell-fate decisions in higher organisms. As dynamical GRN signaling is conserved to humans, this work will facilitate a new understanding of the rules by which life operates. This project will create a new “Sporograph” educational module to teach students how cells make decisions. An experimental module will be based on an inexpensive version of the laboratory B. subtilis optogenetics system developed here, and allow students to induce visible sporulation and biofilm reporter proteins on bacterial agar plates by applying constant or oscillatory light inputs. The Sporograph module will reach thousands of students during this project, and be setup to expand further in future efforts.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.