一种与DNA结合的设计二聚体锌指蛋白的结构。

Structure of a designed dimeric zinc finger protein bound to DNA.

作者信息

Wolfe Scot A, Grant Robert A, Pabo Carl O

机构信息

Department of Biology and the Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.

出版信息

Biochemistry. 2003 Nov 25;42(46):13401-9. doi: 10.1021/bi034830b.

DOI:10.1021/bi034830b

PMID:14621985

Abstract

Proteins that employ dimerization domains to bind cooperatively to DNA have a number of potential advantages over monomers with regards to gene regulation. Using a combination of structure-based design and phage display, a dimeric Cys(2)His(2) zinc finger protein has been created that binds cooperatively to DNA via an attached leucine zipper dimerization domain. This chimera, derived from components of Zif268 and GCN4, displayed excellent DNA-binding specificity, and we now report the 1.5 A resolution cocrystal structure of the Zif268-GCN4 homodimer bound to DNA. This structure shows how phage display has annealed the DNA binding and dimerization domains into a single functional unit. Moreover, this chimera provides a potential platform for the creation heterodimeric zinc finger proteins that can regulate a desired target gene through cooperative DNA recognition.

摘要

在基因调控方面，利用二聚化结构域与DNA协同结合的蛋白质相对于单体具有许多潜在优势。通过基于结构的设计和噬菌体展示相结合的方法，构建了一种二聚化的Cys(2)His(2)锌指蛋白，它通过连接的亮氨酸拉链二聚化结构域与DNA协同结合。这种嵌合体源自Zif268和GCN4的组件，表现出优异的DNA结合特异性，我们现在报道了与DNA结合的Zif268 - GCN4同二聚体的1.5埃分辨率共晶体结构。该结构展示了噬菌体展示如何将DNA结合结构域和二聚化结构域退火成一个单一的功能单元。此外，这种嵌合体为创建异二聚化锌指蛋白提供了一个潜在平台，该蛋白可以通过协同DNA识别来调控所需的靶基因。