FORMINS AND NATIVE COMPLEXES: REGULATION AND FUNCTION

福尔明和天然复合物：调节和功能

• 批准号: 7723632
• 负责人: Bruce L Goode
• 金额: $ 0.81万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7723632
• 关键词: Actins; Affect; Affinity; Animal Model; Architecture; Biochemical Genetics; Cell Polarity; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Environment; Funding; Goals; Grant; Institution; Life; Ligands; Morphogenesis; Myosin ATPase; Pathway interactions; Proteins; Regulation; Research; Research Personnel; Resources; Saccharomyces cerevisiae; Source; United States National Institutes of Health; attenuation; cell growth; in vivo; polarized cell; research study; tandem mass spectrometry

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our goal is to understand how formin activities are regulated in vivo spatially and temporally. Given the potent effects formins have on actin assembly, they must be tightly controlled in the environment of a living cell. Our hypothesis is that through controlled local activation and attenuation of formins, their activities are harnessed to produce actin networks with highly specialized architectures and functions. What are these regulatory mechanisms? We are approaching this question using the model organism S. cerevisiae. The two S. cerevisiae formins Bni1 and Bnr1 assemble sets of highly dynamic actin cables, which in turn direct myosin-dependent polarized cell growth and morphogenesis. To elucidate the pathways controlling Bni1 and Bnr1 activities, we are using a set of affinity tags on known cell polarity factors and formin ligands to isolate the native protein complexes they form, and tandem mass spectrometry to identify the components of these complexes. In parallel, we are performing biochemical and genetic experiments to determine precisely how these proteins affect formin activities and contribute to cell polarity.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们的目标是了解福明活性在体内的空间和时间上是如何受到调节的。鉴于福尔明对肌动蛋白组装的强大影响，必须在活细胞环境中严格控制它们。我们的假设是，通过受控的福尔明局部激活和减弱，它们的活动被利用来产生具有高度专业化结构和功能的肌动蛋白网络。这些监管机制是什么？我们正在使用模式生物酿酒酵母来解决这个问题。两种酿酒酵母形成 Bni1 和 Bnr1 组装多组高动态肌动蛋白电缆，进而指导肌球蛋白依赖性极化细胞生长和形态发生。为了阐明控制 Bni1 和 Bnr1 活性的途径，我们在已知的细胞极性因子和福明配体上使用一组亲和标签来分离它们形成的天然蛋白质复合物，并使用串联质谱法来鉴定这些复合物的成分。与此同时，我们正在进行生化和遗传实验，以精确确定这些蛋白质如何影响福尔明活性并影响细胞极性。