A High-Throughput Screening System for Arabidopsis Transcription Factors and Its Application to Med25-Dependent Transcriptional Regulation

被引:128
作者
Ou, Bin [1 ]
Yin, Kang-Quan [1 ]
Liu, Sai-Nan [1 ]
Yang, Yan [1 ]
Gu, Tren [1 ]
Hui, Jennifer Man Wing [1 ]
Zhang, Li [1 ]
Miao, Jin [1 ]
Kondou, Youichi [2 ]
Matsui, Minami [2 ]
Gu, Hong-Ya [1 ,3 ]
Qu, Li-Jia [1 ,3 ]
机构
[1] Peking Univ, Coll Life Sci, Natl Lab Prot Engn & Plant Genet Engn, Beijing 100871, Peoples R China
[2] RIKEN Yokohama Inst, Plant Sci Ctr, Yokohama, Kanagawa 2300045, Japan
[3] Natl Plant Gene Res Ctr Beijing, Beijing 100101, Peoples R China
基金
中国国家自然科学基金;
关键词
Transcription factors; Arabidopsis; yeast-one-hybrid; yeast-two-hybrid; Med25; PLANT DEFENSIN GENE; PROTEIN INTERACTIONS; FLOWERING-TIME; JASMONATE; ACTIVATION; MEDIATOR; INDUCTION; FAMILY; BOX; EXPRESSION;
D O I
10.1093/mp/ssr002
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The activities of transcription factors (TFs) require interactions with specific DNA sequences and other regulatory proteins. To detect such interactions in Arabidopsis, we developed a high-throughput screening system with a Gateway-compatible Gal4-AD-TF library of 1589 Arabidopsis TFs, which can be easily screened by mating-based yeast-one-hybrid (Y1H) and yeast-two-hybrid (Y2H) methods. The efficiency of the system was validated by examining two well-characterized TF-DNA and TF-protein interactions: the CHE-CCA1 promoter interaction by Y1H and NPR1-TGAs interactions by Y2H. We used this system to identify eight TFs that interact with a Mediator subunit, Med25, a key regulator in JA signaling. We identified five TFs that interacted with the GCC-box cis-element in the promoter of PDF1.2, a downstream gene of Med25. We found that three of these TFs, all from the AP2-EREBP family, interact directly both with Med25 and the GCC-box of PDF1.2, suggesting that Med25 regulates PDF1.2 expression through these three TFs. These results demonstrate that this high-throughput Y1H/Y2H screening system is an efficient tool for studying transcriptional regulation networks in Arabidopsis. This system will be available for other Arabidopsis researchers, and thus it provides a vital resource for the Arabidopsis community.
引用
收藏
页码:546 / 555
页数:10
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