TY  - JOUR
A1  - Lehmeyer, Mona
A1  - Kanofsky, Konstantin
A1  - Hanko, Erik K. R.
A1  - Ahrendt, Sarah
A1  - Wehrs, Maren
A1  - Machens, Fabian
A1  - Hehl, Reinhard
T1  - Functional dissection of a strong and specific microbe-associated molecular pattern-responsive synthetic promoter
JF  - Plant Biotechnology Journal
N2  - Synthetic promoters are important for temporal and spatial gene expression in transgenic plants. To identify novel microbe-associated molecular pattern (MAMP)-responsive cis-regulatory sequences for synthetic promoter design, a combination of bioinformatics and experimental approaches was employed. One cis-sequence was identified which confers strong MAMP-responsive reporter gene activity with low background activity. The 35-bp-long cis-sequence was identified in the promoter of the Arabidopsis thaliana DJ1E gene, a homologue of the human oncogene DJ1. In this study, this cis-sequence is shown to be a tripartite cis-regulatory module (CRM). A synthetic promoter with four copies of the CRM linked to a minimal promoter increases MAMP-responsive reporter gene expression compared to the wild-type DJ1E promoter. The CRM consists of two WT-boxes (GGACTTTT and GGACTTTG) and a variant of the GCC-box (GCCACC), all required for MAMP and salicylic acid (SA) responsivity. Yeast one-hybrid screenings using a transcription factor (TF)-only prey library identified two AP2/ERFs, ORA59 and ERF10, interacting antagonistically with the CRM. ORA59 activates reporter gene activity and requires the consensus core sequence GCCNCC for gene expression activation. ERF10 down-regulates MAMP-responsive gene expression. No TFs interacting with the WT-boxes GGACTTTT and GGACTTTG were selected in yeast onehybrid screenings with the TF-only prey library. In transgenic Arabidopsis, the synthetic promoter confers strong and specific reporter gene activity in response to biotrophs and necrotrophs as well as SA.
KW  - plant-pathogen interaction
KW  - plant synthetic biology
KW  - transient reporter gene assays
KW  - transgenic plants
Y1  - 2016
U6  - https://doi.org/10.1111/pbi.12357
SN  - 1467-7644
SN  - 1467-7652
VL  - 14
SP  - 61
EP  - 71
PB  - Wiley
CY  - Hoboken
ER  -