A strong NF-κB p65 responsive cis-regulatory sequence from Arabidopsis thaliana interacts with WRKY40
Transcription factors from mammals and plants, which play a role in innate immunity, interact with the same microbe-associated molecular pattern (MAMP)-responsive sequences from Arabidopsis thaliana.
The interaction of mouse NF-κB p65 with MAMP-responsive sequences containing the core motif GACTTT of the WT-box was investigated. This revealed one sequence, derived from the promoter of the A. thaliana gene At1g76960, a gene with unknown function, to activate NF-κB p65 dependent reporter gene expression in plant cells very strongly. A bioinformatic analysis predicts three putative NF-κB p65 binding sites in this sequence and all three sites are required for reporter gene activation and binding. The sequence is one of the weakest MAMP-responsive sequences previously isolated, but the introduction of a GCC-box increases its MAMP responsivity in combination with upstream WT-box sequences. Although a bioinformatic analysis of the unmutated cis-sequence only predicts NF-κB p65 binding, A. thaliana WRKY40 was selected in a yeast one-hybrid screen. WRKY40, which is a transcriptional repressor, requires the sequence TTTTCTA for direct binding. This sequence is similar to the WK-box TTTTCCAC, previously shown to interact with tobacco NtWRKY12. In summary, this work supports the similarity in binding site recognition between NF-κB and WRKY factors.
KeywordsArabidopsis thaliana MAMP NF-κB Plant synthetic promoter Transient gene expression analysis WRKY WT-box
We would like to thank Elke Faurie for excellent technical assistance. We are grateful to Christopher Eickhorst and Kazuhiko Namikawa for the gift of mouse mRNA. Great thanks to Miriam Becker for technical assistance when using the french press. This work was supported by the Federal Ministry of Education and Research, Germany (Hochschulpakt 2020).
Author contribution statement
RH and KK conceived and designed research. KK, JR, MS, CJS, and LCA conducted experiments. KK, JR, MS, CJS, LCA, and RH analyzed data. RH wrote the manuscript. All authors read and approved the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest
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