A platform for targeted, post-translational control of protein levels in mammalian cells

用于哺乳动物细胞中蛋白质水平的靶向翻译后控制的平台

• 批准号: 1615562
• 负责人: Laura Segatori
• 金额: $ 69.58万
• 依托单位: William Marsh Rice University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1615562&HistoricalAwards=false
• 关键词: platform; targeted; post; translational; control

Cells physiologic states are dictated by the collective behavior of thousands of molecular components and may vary dramatically depending on environmental factors, resulting in non-linear behaviors. Despite significant advances in systems and synthetic biology, protein function in cells remains hard to decipher due to the plasticity and adaptability of cellular regulatory and quality control mechanisms. The overarching goal of this project is to generate a platform technology to modulate the accumulation of target proteins in cells. This technology is expected to provide exquisite dynamic control over protein expression, thereby providing a tool to study protein function. A series of complementary strategies will be explored to achieve stimulus-dependent control over modulation of protein expression and to deploy this technology to study complex biological processes. As a result, this study is expected to promote progress of fundamental science , advance the bioeconomy, and enable advances in healthcare. By providing broad reaching educational and training opportunities to students belonging to underrepresented groups, this project is also expected to support education and diversity and to benefit society. To overcome the limitations of existing methods for regulating protein steady state levels, the research team built a platform for modulating protein expression post-translationally using a bifunctional system consisting of a molecular recognition unit for target specificity and selectivity, and a degradation-signaling unit that specifies the rate and mechanism of degradation of the target protein. The goal of the proposed research is to engineer this technology to generate a universal platform for controlling protein levels through exogenous or endogenous stimuli. To achieve this goal, three research objectives will be pursued: 1) engineer stimulus-responsive degradation of protein depletion, 2) enhance the dynamic resolution of stimulus-dependent circuits through controlled reporter depletion, and 3) decipher and manipulate the hypoxia signaling network. This research program will be leveraged to provide training, research, and mentoring opportunities for high school teachers and students from the Greater Houston area, and to improve teaching and learning for college students.

细胞的生理状态由数以千计的分子组成的集体行为决定，并可能因环境因素而发生巨大变化，从而导致非线性行为。尽管在系统和合成生物学方面取得了重大进展，但由于细胞调控和质量控制机制的可塑性和适应性，细胞中的蛋白质功能仍然很难破译。该项目的首要目标是产生一种平台技术来调节目标蛋白在细胞中的积累。这项技术有望提供对蛋白质表达的精细动态控制，从而为研究蛋白质功能提供一种工具。将探索一系列补充策略，以实现对蛋白质表达调控的刺激依赖控制，并将这项技术应用于研究复杂的生物过程。因此，这项研究有望促进基础科学的进步，促进生物经济，并使医疗保健方面的进步成为可能。通过向代表不足群体的学生提供广泛的教育和培训机会，该项目还有望支持教育和多样性，并造福社会。为了克服现有蛋白质稳定水平调控方法的局限性，研究小组建立了一个翻译后调控蛋白质表达的平台，该平台使用一个双功能系统，该系统由一个用于靶标特异性和选择性的分子识别单元和一个指定目标蛋白质降解速度和机制的降解信号单元组成。这项拟议研究的目标是设计这项技术，以产生一个通过外源或内源刺激控制蛋白质水平的通用平台。为了实现这一目标，将追求三个研究目标：1)设计蛋白质耗竭的刺激响应降解；2)通过受控的报告耗竭提高刺激依赖电路的动态分辨率；3)破译和操纵缺氧信号网络。这项研究计划将被用来为来自大休斯顿地区的高中教师和学生提供培训、研究和指导机会，并改善大学生的教和学。

项目成果

Zinc Oxide Particles Induce Activation of the Lysosome–Autophagy System

• DOI: 10.1021/acsomega.8b01497
• 发表时间: 2019-01
• 期刊: ACS Omega
• 影响因子: 4.1
• 作者: L. Popp;Laura Segatori

A yeast selection system for the detection of proteasomal activation

• DOI: 10.1093/protein/gzz006
• 发表时间: 2018-11-01
• 期刊: PROTEIN ENGINEERING DESIGN & SELECTION
• 影响因子: 2.4
• 作者: Zhao，Wenting;Bachhav，Bhagyashree;Segatori，Laura
• 通讯作者: Segatori，Laura

Autophagic response to cellular exposure to titanium dioxide nanoparticles

• DOI: 10.1016/j.actbio.2018.08.021
• 发表时间: 2018-10-01
• 期刊: ACTA BIOMATERIALIA
• 影响因子: 9.7
• 作者: Popp, Lauren;Vinh Tran;Segatori, Laura
• 通讯作者: Segatori, Laura