Functional dissection of a strong and specific microbe-associated molecular pattern-responsive synthetic promoter
| Autor: | Maren Wehrs, Fabian Machens, Reinhard Hehl, Mona Lehmeyer, Sarah Ahrendt, Konstantin Kanofsky, Erik K. R. Hanko |
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| Rok vydání: | 2015 |
| Předmět: |
0106 biological sciences
0301 basic medicine DNA Plant Transgene Arabidopsis Plant Science Genes Plant 01 natural sciences 03 medical and health sciences Gene Expression Regulation Plant Genes Reporter Gene expression Gene Regulatory Networks Promoter Regions Genetic Gene Transcription factor Institut für Biochemie und Biologie Genetics Reporter gene biology Pathogen-Associated Molecular Pattern Molecules fungi Promoter Plants Genetically Modified biology.organism_classification 030104 developmental biology Bioreporter Salicylic Acid Agronomy and Crop Science Transcription Factors 010606 plant biology & botany Biotechnology |
| Zdroj: | Plant Biotechnology Journal. 14:61-71 |
| ISSN: | 1467-7644 |
| DOI: | 10.1111/pbi.12357 |
| Popis: | Summary 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 one-hybrid 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. |
| Databáze: | OpenAIRE |
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