The Development of Protein Microarrays and Their Applications in DNA-Protein and Protein-Protein Interaction Analyses of Arabidopsis Transcription Factors

作     者：Wei Gong Kun He Mike Covington S. R Dinesh-Kumar Michael Snyder Stacey L. Harmer Yu-Xian Zhu Xing Wang Denga 

作者机构：Peking-Yale Joint Center for Plant Molecular Genetics and Agrobiotechnology Co and Plant Genetic Engineering Peking University Beijing 100871 China Department of Molecular Cellular and Developmental Biology Yale University New Haven Connecticut 06520 USA Section of Plant Biology College of Biological Sciences University of California Davis 95616 USA 

出 版 物：《Molecular Plant》 (分子植物（英文版）)

年 卷 期：2008年第1卷第1期

页      面：27-41页

核心收录：

学科分类：0710[理学-生物学] 071001[理学-植物学] 07[理学] 0901[农学-作物学] 0902[农学-园艺学] 

基　　金：We thank William Terzaghi for reading and commenting on this manuscript. This work was supported by the Chinese National Natural Science Foundation and in part by The National Science Foundation 2010 Program (US) and by the National Institutes of Health grant number GM069418 to SLH 

主　　题：transcription factor (TF) TF-DNA EE (evening element) 

摘      要：We used our collection of Arabidopsis transcription factor （TF） ORFeome clones to construct protein microarrays containing as many as 802 TF proteins. These protein microarrays were used for both protein-DNA and proteinprotein interaction analyses. For protein-DNA interaction studies, we examined AP2/ERF family TFs and their cognate cis-elements. By careful comparison of the DNA-binding specificity of 13 TFs on the protein microarray with previous non-microarray data, we showed that protein microarrays provide an efficient and high throughput tool for genome-wide analysis of TF-DNA interactions. This microarray protein-DNA interaction analysis allowed us to derive a comprehensive view of DNA-binding profiles of AP2/ERF family proteins in Arabidopsis. It also revealed four TFs that bound the EE （evening element） and had the expected phased gene expression under clock-regulation, thus providing a basis for further functional analysis of their roles in clock regulation of gene expression. We also developed procedures for detecting protein interactions using this TF protein microarray and discovered four novel partners that interact with HY5, which can be validated by yeast two-hybrid assays. Thus, plant TF protein microarrays offer an attractive high-throughput alternative to traditional techniques for TF functional characterization on a global scale.