Genetic Dissection of the Specificity of Leucine Zipper Dimerization

所提出的研究应用经典的噬菌体遗传方法， 大规模组合一级结构文库的筛选 与酵母转录因子GCN 4的亮氨酸拉链相关。 这些研究的目的是定义可以形成的序列 具有野生型GCN 4亮氨酸拉链的异二聚体，以及 表征异源二聚化的一级序列要求。 %%% 氨基酸侧链在蛋白质二聚化中的作用 将通过改变主要顺序来评估过程 随机的简单亮氨酸拉链结构基序。 的 每个新结构的功能意义（将有 数百万个这样的结构）将被筛选，在一个基于遗传学的 系统，在二聚化的能力方面。 两类人群 将被测序：（1）那些可以形成异二聚体的分子 与野生型GCN 4亮氨酸拉链，和（2）这些分子 它们形成稳定的同源二聚体，但不形成异源二聚体（与野生的- 型亮氨酸拉链）。 这样的研究会让我们 进入肽-肽的结构限制， 蛋白质相互作用

The proposed studies apply classical phage genetic approaches to the screening of massive combinatorial primary structure libraries related to the leucine zipper of yeast transcription factor GCN4. The object of these studies is to define sequences which can form heterodimers with wild type GCN4 leucine zippers, and to characterize primary sequence requirements for heterodimerization. %%% The roles of amino acid side chains in protein dimerization processes will be evaluated by altering the primary sequence randomly of a simple leucine zipper structural motif. The functional significance of each new structure (there will be millions of such structures) will be screened, in a genetics based system, in terms of ability to dimerize. Two types of populations will be sequenced: (1) those molecules which can form heterodimers with the wild-type GCN4 leucine zipper, and (2) those molecules which form stable homodimers, but not heterodimers (with the wild- type leucine zipper). Such studies will shed considerable insight into the structural constraints involved in peptide-peptide, protein-protein interactions.