FORMINS AND NATIVE COMPLEXES: REGULATION AND FUNCTION

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 中心，不一定是研究者的机构。 我们的目标是要了解如何在体内的空间和时间调节的生物活性。鉴于formins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，通过控制局部激活和衰减的formin，他们的活动被利用，产生肌动蛋白网络高度专业化的架构和功能。这些监管机制是什么？我们正在用模式生物S来探讨这个问题。啤酒。两个S。酿酒酵母形成蛋白Bni 1和Bnr 1组装成一组高度动态的肌动蛋白电缆，这反过来又指导肌球蛋白依赖的极化细胞生长和形态发生。为了阐明控制Bni 1和Bnr 1活性的途径，我们正在使用一组已知细胞极性因子和Bnr 1配体上的亲和标签来分离它们形成的天然蛋白质复合物，并使用串联质谱来鉴定这些复合物的组分。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细胞活性并有助于细胞极性。

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.