FORMINS AND NATIVE COMPLEXES: REGULATION AND FUNCTION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们的目标是了解福尔马林活性在体内是如何在空间和时间上受到调控的。鉴于福尔马林对肌动蛋白组装的强大影响，它们必须在活细胞的环境中受到严格控制。我们的假设是，通过受控的局部激活和福尔马林的衰减，它们的活性被利用来产生具有高度专业化的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。两个酿酒酵母形成蛋白Bni1和Bnr1组装了一组高度动态的肌动蛋白电缆，这些电缆反过来指导肌球蛋白依赖的极化细胞的生长和形态发生。为了阐明控制Bni1和Bnr1活性的途径，我们正在使用一组已知细胞极性因子和Form配体上的亲和标记来分离它们形成的天然蛋白质复合体，并用串联质谱仪来鉴定这些复合体的组成。与此同时，我们正在进行生化和遗传学实验，以准确确定这些蛋白质如何影响Forin活性并对细胞极性做出贡献。

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.