Tracking shallow and dynamic chemoattractant gradients - how yeast cells amplify both internal and external signals to locate mating partners

跟踪浅层和动态趋化剂梯度——酵母细胞如何放大内部和外部信号来定位交配伙伴

• 批准号: 2341919
• 负责人: David Stone
• 金额: $ 162.62万
• 依托单位: University of Illinois at Chicago
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-03-01 至 2028-02-29
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2341919&HistoricalAwards=false
• 关键词: Tracking; shallow; dynamic; chemoattractant; gradients

This project will contribute to our understanding of gradient sensing, the ability of cells to sense small differences in chemical concentration across their surfaces, and thereby locate the source of the stimulus. This phenomenon is essential for the development and health of all organisms. The PI uses yeast cells as a model to study the molecular mechanisms underlying gradient sensing, which are thought to be broadly applicable to cells in more complex organisms. In previous studies, he discovered a “gradient tracking machine” that cells must assemble before they are able to sense the direction of the chemical source. In this investigation, he will continue to investigate how this machine functions to decode chemical gradients. During this project period, the PI and his senior research specialist will mentor select biology students from the City Colleges of Chicago, to enhance their chances of graduating from a four-year institution with a BS in a STEM field. The proposed undergraduate research and mentoring program is designed to inspire, instruct, advise, and support underrepresented students who are interested in a STEM career. Two outstanding candidates will be chosen to participate each year based on their academic potential and motivation to pursue a STEM major at a four-year institution. By participating in regular tutoring sessions, paid summer research internships in the PI’s lab, and public outreach events, students will gain experience conducting scientific research, an ability to critically evaluate the research of others, and practice presenting their work. This program is expected to increase the chances of eight students to succeed as science majors at four-year institutions. The investigation will also provide the PI's students with interdisciplinary training through interactions with collaborators who are experts in diverse areas.The best-known gradient-stimulated cellular outputs, chemotaxis (directed cell movement), and chemotropism (directed cell growth), are required for a wide range of biological processes. Although they ultimately exhibit quite different behavior, chemotactic and chemotropic cells face similar challenges: the responding cell must sense small differences in chemical concentration across its surface, determine the direction of the gradient source, and polarize its cytoskeleton toward it. The mating response of the budding yeast S. cerevisiae is chemotropic: mating cells interpret complex pheromone gradients and polarize their growth in the direction of the closest partner. Like many chemosensing cells, yeasts use G protein-coupled receptors to detect chemoattractant. The goal of this project is to understand how yeast cells accurately sense direction in shallow, complex, and dynamic pheromone gradients. Based on discoveries made in a previous project, the PI published a deterministic model of gradient sensing that explains, in broad terms, how yeast cells translate a vanishingly small differential of activated receptors across their surfaces into accurate and robust directional responses. Mating yeast initially ignore the pheromone gradient, as they first colocalize signaling, polarity, and trafficking proteins to the default polarity site they use for budding, building a “gradient tracking machine” (GTM). Once assembled, the GTM moves along the plasma membrane to the point of maximal pheromone concentration, where it marks the chemotropic site used for mating. The primary negative regulator of G-protein signaling, the RGS protein Sst2, is essential for this process, and phosphorylation of one of the G-protein subunits (Gbetagamma)plays a critical role. The priorities of this investigation are to learn how Sst2 is controlled in space and time, how the phosphorylation of the G protein subunit contributes to gradient tracking, and how dynamic intercommunication between GTMs enables mating partners to orient toward a common fusion site. These questions will be answered using imaging, genetic, biochemical, proteomic, and computational approaches.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.

这个项目将有助于我们对梯度感应的理解，即细胞感知其表面化学物质浓度的微小差异的能力，从而定位刺激的来源。这种现象对所有生物的发育和健康都是必不可少的。PI使用酵母细胞作为模型来研究梯度感应的分子机制，这被认为广泛适用于更复杂生物体中的细胞。在之前的研究中，他发现了一种“梯度跟踪机器”，细胞必须在它们能够感知化学源的方向之前组装起来。在这次调查中，他将继续研究这台机器如何解码化学梯度。在这个项目期间，PI和他的高级研究专家将指导从芝加哥城市学院挑选的生物学学生，以提高他们从四年制大学毕业并获得STEM领域学士学位的机会。拟议的本科研究和指导计划旨在激励、指导、建议和支持对STEM职业感兴趣的代表性不足的学生。每年将根据他们的学术潜力和在四年制大学攻读STEM专业的动机，选择两名优秀的候选人参加。通过参加定期的辅导课程，在PI实验室的带薪暑期研究实习，以及公共宣传活动，学生将获得进行科学研究的经验，批判性地评估他人研究的能力，并练习展示自己的工作。预计该项目将增加8名学生进入4年制大学理科专业的机会。调查还将通过与不同领域的专家合作，为PI的学生提供跨学科的培训。最著名的梯度刺激细胞输出，趋化性（定向细胞运动）和趋化性（定向细胞生长），是广泛的生物过程所必需的。尽管趋化细胞和趋化细胞最终表现出截然不同的行为，但它们面临着相似的挑战：响应细胞必须感知其表面化学浓度的微小差异，确定梯度源的方向，并将其细胞骨架向其极化。出芽酵母的交配反应是趋化的：交配细胞解释复杂的信息素梯度，并向最接近的伴侣方向极化生长。像许多化学感应细胞一样，酵母使用G蛋白偶联受体来检测化学引诱物。这个项目的目标是了解酵母细胞如何在浅的、复杂的和动态的信息素梯度中准确地感知方向。基于先前项目的发现，PI发表了一个梯度感应的确定性模型，从广义上解释了酵母细胞如何将其表面上微小的激活受体差异转化为准确而稳健的定向反应。交配酵母最初忽略了信息素梯度，因为它们首先将信号、极性和蛋白质运输到它们用于出芽的默认极性位置，从而建立了一个“梯度跟踪机器”（GTM）。一旦组装，GTM沿着质膜移动到信息素浓度最大的点，在那里它标记了用于交配的趋化位点。g蛋白信号传导的主要负调控因子RGS蛋白Sst2在这一过程中至关重要，其中一个g蛋白亚基（Gbetagamma）的磷酸化起着关键作用。这项研究的重点是了解Sst2如何在空间和时间上受到控制，G蛋白亚基的磷酸化如何促进梯度跟踪，以及GTMs之间的动态相互通信如何使交配伙伴朝向共同的融合位点。这些问题将回答使用成像，遗传，生化，蛋白质组学和计算方法。该奖项反映了美国国家科学基金会的法定使命，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。