拟南芥AtHHR2与DREB2C的相互作用研究

Analysis of the interaction between AtHHR2 and DREB2C in Arabidopsis thaliana

摘要:

已有研究表明拟南芥Highly Homologous RING domain 2（At5g43200，命名为AtHHR2）基因在盐胁迫中起正调控作用，并且其同源基因AtHHR3基因在酵母双杂交实验筛选中与DREB2C基因有相互作用.本研究利用体外、体内蛋白质相互作用实验以及生物化学方法进一步探究了AtHHR2的功能性.体外Pulldown实验证实了AtHHR2与DREB2C有体外的相互作用.体外泛素化实验进一步验证了它们之间的相互作用，且AtHHR2起到E3连接酶的作用.为了进一步确定它们的关系，我们用双分子荧光互补实验，在体内分别验证了AtHHR2与DREB2C之间确实有相互作用.综上所述在拟南芥中AtHHR2与DREB2C有相互作用关系，且AtHHR2起到E3连接酶的作用.

Abstract:

A previous report has revealed that AtHHR2 (At5g43200) played a positive regulatory role in Arabidopsis thaliana in salt stress, and its homologous gene AtHHR3 has interaction with DREB2C gene in yeast twohybrid test. In this study, in vitro and in vivo protein interaction assays and biochemical methods were used to further explore the functionality of AtHHR2. And Pulldown result validated the interaction between AtHHR2 and DREB2C in vitro. And the ubiquitination analysis demonstrated that AtHHR2 acted as an E3 ligase of DREB2C, further confirming the interaction between AtHHR2 and DREB2C. Furthermore, bimolecular fluorescence complementation assay results showed that AtHHR2 did have interaction with DREB2C in vivo. In summary, the AtHHR2 interacts with DREB2C and acts as an E3 ligase.