转录因子ZmMYB153与TPL/TPRs蛋白的互作研究

Study of interaction between transcription factor ZmMYB153 and TPL/TPRs proteins

转录因子MYB44通过EAR基序与TPL/TPRs蛋白直接互作,参与植物抵抗生物和非生物胁迫过程,其同源蛋白ZmMYB153的功能和调控机制尚未明确。本研究利用酵母双杂交(Yeast two-hybrid,Y2H)技术,对ZmMYB153与TPL/TPRs(TOPLESS/TOPLESS-related proteins)蛋白之间的互作关系进行研究,结果发现,共同转化ZmMYB153与TPR2组合(AD-ZmMYB153+BD-TPR2、BD-ZmMYB153+AD-TPR2)的酵母菌落在-Leu/-Trp、-Leu/-Trp/-His和-Leu/-Trp/-His+3-AT培养基上均能正常生长;而其他组合以及阴性对照组合的酵母菌落均不能在-Leu/-Trp/-His+3-AT培养基上正常生长,表明ZmMYB153与TPR2蛋白在酵母细胞中能够直接互作;与阳性对照相比,共转化EAR基序突变的ZmMYB153-mEAR与TPR2组合(AD-ZmMYB153-mEAR+BD-TPR2、BD-ZmMYB153-mEAR+AD-TPR2)的酵母不能在-Leu/-Trp/-His+3-AT培养基上生长,表明EAR基序是ZmMYB153与TPR2蛋白在酵母中互作的关键位点。研究结果为阐明ZmMYB153在玉米生长发育和抵抗生物/非生物胁迫中的功能及其调控机制奠定了基础。

Transcription factor MYB44 interacts directly with TPL/TPRs proteins through the EAR motif and participates in plant resistance to biotic and abiotic stress.The function and regulatory mechanism of its homologous protein ZmMYB153 has not yet been clarified.This study uses yeast two-hybrid(Y2 H)technology to study the interaction between ZmMYB153 and TPL/TPRs(TOPLESS/TOPLESS-related proteins).The results showed that the yeast colonies that co-transformed with ZmMYB153 and TPR2(AD-ZmMYB153+BD-TPR2,BDZmMYB153+AD-TPR2)could grow normally on medium of-Leu/-Trp,-Leu/-Trp/-His,and-Leu/-Trp/-His+3-AT.However,the yeast colonies of the other combinations and the negative control combination could not grow normally on the-Leu/-Trp/-His+3-AT medium,indicating that ZmMYB153 and TPR2 proteins can directly interact in yeast cells.Compared with the positive control,the yeast that co-transformed with the combination of ZmMYB153-mEAR and TPR2 mutation(AD-ZmMYB153-mEAR+BD-TPR2,BD-ZmMYB153-mEAR+AD-TPR2)could not be found in-Leu/-Trp/-His+3-AT medium,indicating that the EAR motif is a key site for the interaction of ZmMYB153 and TPR2 protein in yeast.The results lay the foundation for elucidating the function of ZmMYB153 in maize growth,development and resistance to biotic and abiotic stress as well as its regulatory mechanism.