一种大豆LEA4蛋白PM1与AE7/MIP18相互作用对植物细胞质铁硫簇组装系统CIA功能的影响

植物种子发育晚期将经历一个脱水阶段，LEA家族成员在此阶段表达富集，参与保护细胞中生物大分子。对生物体功能有重要作用的铁硫蛋白，其组装复合体成员的缺失往往导致植株胚胎致死，因此对其分子活性的保护是至关重要的。在前期研究中，我们利用酵母双杂交系统筛选并证明了大豆PM1蛋白与大豆细胞质铁硫簇组装系统复合体成员AE7/MIP18间存在相互作用。本项目拟在此基础之上，根据PM1蛋白自身的序列特征，进一步采用酵母双杂交、Pull Down、双分子荧光互补及免疫共沉淀实验等技术，研究PM1与AE7/MIP18间的相互作用关系，并通过检测转基因植株对于DNA损伤的耐受性来验证CIA复合体的功能，证明PM1蛋白对于CIA复合体功能的影响。从蛋白质间相互作用的角度阐释LEA蛋白在种子发育脱水阶段对生物分子的保护功能及其分子机理，进一步揭示固有无序蛋白在植物逆境应答中的功能，为作物抗逆分子育种提供理论依据。

Plant late embryogenesis abundant (LEA) proteins will highly express and accumulate at the late stage during seeds developing, and protect the biomolecule while the dehydration process. Iron-sulfur protein, a protein family which each contains a Fe-S cluster center, plays an important role in many metabolism pathways. Lost function of members in the Fe-S cluster assembly system will cause plants embryonic death. We have found that soybean LEA protein PM1 interacted with cytoplasmic iron-sulfur cluster assembly (CIA) system protein AE7/MIP18 using yeast two-hybrid (Y2H) experiment. Base on this result, we will analyze the relationship between PM1 and protein members of CIA complex by Y2H, pull down, BiFC and Co-IP experiments. Use tolerance to DNA damage as a phenotype for structural and functional integrity of CIA complex. Try to reveal the protective function of LEA protein in dehydration stage of seed developing from the angle of protein-protein interaction. Enrich the knowledge about the functions of intrinsically disordered proteins during stress resistance.