CAREER: Molecular mechanisms underlying yeast cellular starvation tolerance; spatial reasoning to increase STEM participation

所有生物都必须对环境的变化作出反应。例如，生长在葡萄上的酵母必须在干旱、突然下雨和发酵过程中酒精浓度的变化中存活。为了应对压力，细胞会改变其内部的组成和化学性质。一个突出的问题是细胞内部的变化如何导致对压力的保护。核磁共振光谱（NMR）是一种可以提供细胞内蛋白质原子水平行为信息的技术。在这项研究中，NMR将用于测量细胞受到压力时蛋白质行为的变化。此外，该项目将促进对小学教师及其学生进行空间推理方面的培训。空间推理被广泛地定义为视觉化、心理操纵和创建复杂三维物体的表示的能力。一个小规模的文化误解是，空间推理技能是不变的;事实上，它们是最可训练的STEM相关技能之一。通过针对小学生，该项目将有助于提高STEM的总体参与率，并减少代表性不足群体在STEM方面的差距。细胞通过细胞内化学环境、蛋白质定位和动力学以及整体细胞质物理性质的大范围变化来响应环境扰动。所有这些反应都被认为是细胞存活所必需的。为了协调应激反应蛋白的敏感性与应激条件下细胞环境中的未知变化，我们需要细胞内蛋白行为的原子尺度读数。在本研究中，细胞内核磁共振实验将被用来探测芽殖酵母的细胞内环境。芽殖酵母具有良好的遗传、环境和药理学扰动，可用于选择性地控制细胞应激反应的各个方面。通过NMR比较溶液和细胞蛋白质的行为将用于确定蛋白质在细胞中如何变化以及细胞应激如何进一步影响蛋白质。NMR将用于映射应激反应蛋白质的原子水平变化，蛋白质片段库将用于确定应激反应的序列特异性。酵母细胞的操作将用于确定导致这些变化的潜在化学扰动。该奖项反映了NSF的法定使命，并被认为值得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。

All organisms must respond to changes in their environment. For example, yeast which grow on grapes must survive drought, sudden rain, and changes in alcohol concentration during fermentation. In response to stress, cells alter the composition and chemical properties of their interior. An outstanding question is how changes in the interior of cells result in protection from stresses. Nuclear magnetic resonance spectroscopy (NMR) is a technique which can give information on the atomic level behavior of proteins within cells. In this research, NMR will be used to measure changes in protein behavior as cells are stressed. In addition, this project will facilitate the training of elementary school teachers and their students in spatial reasoning. Spatial reasoning is broadly defined as the ability to visualize, mentally manipulate, and create representations of complex three- dimensional objects. A widescale cultural misconception is that spatial reasoning skills are immutable; in fact, they are one of the most trainable of STEM-related skills. By targeting elementary school students, this project will serve to increase STEM participation in general and to mitigate disparities in STEM for underrepresented groups. Cells respond to environmental perturbations through wide-scale changes in their intracellular chemical environment, protein localization and dynamics, and global cytoplasmic physical properties. All of these responses are thought to be necessary for cell survival. To reconcile the sensitivity of stress-responsive proteins with the unknown changes in the cellular environment under stress conditions, we require an atomic-scale readout of intracellular protein behavior. In this research, in cell NMR experiments will be used to probe the intracellular environment in budding yeast. Budding yeast have well developed genetic, environmental and pharmacological perturbations that can be used to selectively control aspects of the cellular stress response. Comparisons between solution and cellular protein behavior by NMR will be used to determine how proteins change in cells and how cellular stress further effects proteins. NMR will be used to map atomic-level changes in stress-responsive proteins and a library of protein fragments will be used to determine sequence specificity of stress response. Manipulations of the yeast cells will be used to determine underlying chemical perturbations that lead to those changes.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.