Screening and validation of candidate proteins interacting with TR1 in Arabidopsis thaliana

Abstract:

TR1 is an E3 ligase localized in the plasma membrane, and the previous studies indicated that it played a key role in salt, drought and heat stress response in many plants.However, the molecular mechanism of its function is unclear, and the target protein interacting with it is not found.In this study, a yeast two-hybrid system screening using TR1-△119 as a bait was performed to screen the Normalized Universal Arabidopsis cDNA library and found 13 candidate proteins with potential interaction. Forward and reverse two-hybrid system multiple validation revealed that diploid yeast containing TR1 and CURT1C or TR1 and SWEET5 will grow on SD/-Trp/-Leu/-His and SD/-Trp/-Leu/-His/-Ade/X-α-gal minimal media and see blue colonies. BiFC technique also demonstrated that TR1 interacts with CURT1C and SWEET5,providing the foundation for further understanding of the functions and molecular reaction tolerance mechanism．