水稻锌指蛋白ZFP36互作蛋白的筛选及互作蛋白基因表达

Screening the Interacting Protein of ZFP36 and the Expression of the Interacting Protein in Oryza sativa

C_(2)H_(2)类型的锌指蛋白转录因子在植物生长发育和应对逆境胁迫中发挥重要作用。水稻(Oryza sativa L.)中,一种C_(2)H_(2)类型的锌指蛋白ZFP36(zinc finger protein 36)是脱落酸(abscisic acid,ABA)介导的细胞信号转导中的重要组分。为了研究水稻ZFP36在ABA介导的细胞信号转导中作用机制,本研究采用酵母双杂交系统,将ZFP36作为诱饵蛋白,通过筛选经ABA诱导的水稻叶片获得的酵母文库,筛选出59个阳性克隆。进一步通过酵母双杂交(yeast two-hybrid system,Y2H)和萤火虫荧光素酶互补成像(firefly luciferase complementary imaging,LCI)系统,最终获得4个与ZFP36互作的蛋白,包括OsAE7(asymmetric leaves1/2 enhancer 7)、OsDjC46(heat shock protein)、OsDIP1(R3H domain containing protein)和OsLEA5(late embryogenesis abundant protein)。基于实时荧光定量PCR(quantitative real-time PCR,RT-qPCR)分析,结果表明编码这四个蛋白的基因均响应ABA及非生物胁迫的诱导。该研究可为进一步阐明ZFP36在ABA介导的细胞信号转导过程中的分子机制提供科学参考。

The transcription factor of C_(2)H_(2)-type zinc finger protein plays a key role in plant growth and development,as well as under adverse environment.ZFP36 is one of the C_(2)H_(2)-type zinc finger protein in rice,which plays a pivotal role in abscisic acid(ABA)-mediated signal transduction.In order to investigate the detail molecular mechanism of ZFP36 to modulate ABA-mediated antioxidant defence,here,we used the full-length of ZFP36 as a bait to screen the cDNA library from abscisic acid(ABA)-treated rice leaf by yeast two-hybrid screening system,and 59 positive clones were selected.Further,4 interacting proteins of ZFP36 were identified via Y2 H method and firefly luciferase complementation imaging(LCI)system,including asymmetric leaves1/2 enhancer 7(OsAE7),heat shock protein(OsDjC46),R3 H domain-containing protein(OsDIP1),and late embryogenesis abundant protein(OsLEA5).Moreover,we found that all of the genes coding the 4 proteins were induced by ABA and other abiotic stress treatment via quantitative real-time PCR(RT-qPCR)analysis.This study will be conductive to illustrate the molecular mechanism of ZFP36 in ABA-signaling.