RRC ID 54368
Author 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.
Title A high-throughput screening system for Arabidopsis transcription factors and its application to Med25-dependent transcriptional regulation.
Journal 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.
Volume 4(3)
Pages 546-55
Published 2011-5
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 10.812
Times Cited 91
Resource
Arabidopsis / Cultured plant cells, genes pda