PHEROMONE INDUCED CELL POLARITY IN S CEREVISIAE

酿酒酵母将用于研究细胞极化， 响应外部信号。细胞极化是一个基本的 生物现象。基础过程，例如不对称单元 分裂、趋化性和神经突生长。in s.酿酒厂 单倍体a和α细胞向交配源生长 为交配做准备的信息素极性建立 蛋白质被定位于新生长的位点，并将 细胞骨架朝向该位点。极性建立蛋白质 被招募到生长部位，以及它们 对细胞骨架的影响尚不清楚。我将研究 这些蛋白质以及它们如何通过最初聚焦于 BEM1是一个极性建立基因。BEM1-绿色荧光蛋白 (GFP)融合将用于确定BEM1的定位动力学。 BEM1的结构-功能分析将定义重要的蛋白质结构域 为了它的本地化。将使用两种方法来确定组件 对于BEM1功能重要的是：1）双混合方法将识别 与BEM1发生物理相互作用的蛋白质;这些蛋白质可以起以下作用： BEM1的上游或下游。2)将进行基因筛查， 鉴定参与极性建立基因定位的基因座。 这些基因座中的突变体将无法与bem 1交配（基于Chenevert。 等人[（1994）Genetics，136：1287 - 1297]），并且不能正确定位 BEM1-GFP。

Saccharomyces cerevisiae will be used to study cell polarization in response to an external signal. Cell polarization is a fundamental biological phenomenon. underlying process such as asymmetric cell division, chemotaxis, and neurite outgrowth. In S. cerevisiae, the haploid a and alpha cells polarize and grow toward the source of mating pheromone in preparation for the mating event. The polarity establishment proteins are localized to the site of new growth and orient the cytoskeleton toward this site. How the polarity establishment proteins are recruited to the site of growth and the mechanism by which they affect the cytoskeleton is unknown. I will study the localization of these proteins and how they orient the cytoskeleton by initially focusing on BEM1, a polarity establishment gene. A BEM1-Green Fluorescent Protein (GFP) fusion will be used to determine BEM1's localization kinetics. Structure-function analysis of BEM1 will define protein domains important for its localization. Two approaches will be used to identify components important for BEM1 function: 1) The two-hybrid approach will identify proteins that physically interact with BEM1; these can act either upstream or downstream of BEM1. 2) A genetic screen will be done to identify loci involved in localizing the polarity establishment genes. Mutants in these loci will be unable to mate to bem1 (based on Chenevert. et al. [(1994) Genetics, 136: 1287-1297]) and unable to properly localize BEM1-GFP.