RRC ID |
54368
|
著者 |
Ou B, Yin KQ, Liu SN, Yang Y, Gu T, Wing Hui JM, Zhang L, Miao J, Kondou Y, Matsui M, Gu HY, Qu LJ.
|
タイトル |
A high-throughput screening system for Arabidopsis transcription factors and its application to Med25-dependent transcriptional regulation.
|
ジャーナル |
Mol Plant
|
Abstract |
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.
|
巻・号 |
4(3)
|
ページ |
546-55
|
公開日 |
2011-5-1
|
DOI |
10.1093/mp/ssr002
|
PII |
S1674-2052(14)60586-4
|
PMID |
21343311
|
MeSH |
Arabidopsis / genetics*
Arabidopsis / metabolism*
Arabidopsis Proteins / metabolism*
DNA-Binding Proteins
Gene Expression Regulation, Plant*
Gene Library
High-Throughput Screening Assays / methods*
Models, Biological
Nuclear Proteins / metabolism*
Promoter Regions, Genetic / genetics
Protein Binding
Reproducibility of Results
Transcription Factors / metabolism*
Transcription, Genetic*
|
IF |
12.084
|
引用数 |
91
|
リソース情報 |
シロイヌナズナ / 植物培養細胞・遺伝子 |
pda |