Finding Molecular Targets for Engineering Auxin Signaling with Synthetic Interaction Domains
International Conference on Plant Synthetic Biology and Bioengineering
2021
5th International Conference on Plant Synthetic Biology, Bioengineering and Biotechnology
Poster Session
Poster Session
To untangle this complex regulatory pathway, we designed synthetic ARF and AUX/IAA constructs swapping the nonspecific PB1 interaction domains with highly specific interaction domains of tunable affinity using modular cloning methods. We expressed combinations of these constructs in yeast containing an auxin responsive promoter and TIR1/AFB auxin receptor to determine the impact of protein-protein binding specificity and affinity on gene expression dynamics. Steady-state activation assays performed on strains with only synthetic ARFs and an auxin responsive promoter showed elevated reporter activation which confirms that these synthetic transcription factors are functional in vivo. Strains containing the full auxin response circuit showed an auxin response similar to the native yeast ARC. Comparison of gene expression data between our ARF-Aux/IAA synthetic pairs vs. native pairs will allow us to refine our understanding of the mechanism by which auxin regulates numerous genes.