The N-terminal MYB domains affect the stability and folding aspects of Arabidopsis thaliana MYB4 transcription factor under thermal stress.

Abstract

The MYB4 transcription factor, a member of R2R3-type subfamily of MYB domain protein, plays a key role in the regulation of accumulation of UV-B absorbing phenylpropanoids in Arabidopsis. Although UV-B and thermal stress generate some common stress response, the effect of elevated temperature on the conformational stability of MYB4 remains limited. This study describes the folding and aggregation properties of Arabidopsis MYB4 protein under thermal stress condition. Circular dichroism spectral studies and Bis-ANS binding assays have indicated that the removal of the N-terminal MYB domain affects the structural conformation of the protein and disrupts surface hydrophobic binding sites at higher temperature. Urea-induced equilibrium unfolding studies revealed that the removal of the N-terminal region lowers the thermodynamic stability of MYB4 at elevated temperature. Tryptophan fluorescence spectral pattern and both in vitro and in vivo aggregation studies have revealed the importance of the N-terminal second MYB domain encompassing the N-terminal 62-116 amino acid residues in regulating MYB4 protein stability at higher temperature. On the other hand, comparison of the growth response of wild-type Arabidopsis and atmyb4 mutant line have suggested that MYB4 may not directly affect plant response under thermal stress condition and only marginal role of MYB4 in controlling thermomorphogenesis in Arabidopsis. Interestingly, immunoprecipitation studies have revealed that HSP90 specifically interacts with MYB4 in vivo at the endogenous level, indicating the possible role of HSP90 in governing the stability of MYB4 at elevated temperature in Arabidopsis.