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细胞壁连接的类受体激酶(wall-associated kinase,WAK)是植物细胞中一类特有的类受体激酶基因亚家族,因其胞外域与细胞壁紧密相连而得名.水稻中共有125个OsWAK基因,OsWAK50编码的蛋白质具有胞外域、跨膜域和激酶域,呈现典型的WAK样受体激酶特征.首先通过对OsWAK50-GFP融合蛋白的观察发现OsWAK50定位于细胞膜并且与细胞壁偶联.进而通过酵母双杂交系统筛选到了20个可能与OsWAK50胞内域相互作用的候选蛋白,并通过一对一酵母转化验证了OsSK4、OsSWIB和OsSWI3C全长均可与OsWAK50胞内域相互作用.进一步分析显示,OsSWIB能够直接与OsWAK50激酶域互作,而OsSK4和OsSWI3C与OsWAK50胞内域的互作是依赖于OsWAK50 C端的.研究还表明,OsSK4和OsSWIB亦能与OsWAK50同源基因OsWAK53a结合,而OsSWI3C则不能与OsWAK53a结合.双分子荧光互补实验证明,OsSK4与OsWAK50和OsWAK53a能够在植物体内发生互作.以上结果为阐明OsWAK50发挥功能的分子机制提供了重要线索.
Wall-associated kinase (WAK), a unique receptor-like kinase subfamily of plant cells, is named after its extracellular domain is closely linked to the cell wall. There are 125 OsWAK genes in rice , The protein encoded by OsWAK50 has extracellular domain, transmembrane domain and kinase domain, presenting typical WAK-like receptor kinase characteristics.Firstly, through the observation of OsWAK50-GFP fusion protein, OsWAK50 was localized to the cell membrane and coupled to the cell wall, Two hybrids screened 20 candidate proteins that may interact with the OsWAK50 intracellular domain and verified that the full length OsSK4, OsSWIB and OsSWI3C all interacted with the OsWAK50 intracellular domain through one-to-one yeast transformation.Further analysis showed that OsSWIB Can interact directly with the OsWAK50 kinase domain and the interaction between OsSK4 and OsSWI3C and the OsWAK50 intracellular domain is dependent on the C-terminal OsWAK 50. The study also showed that OsSK4 and OsSWIB also bind to the OsWAK50 homologue OsWAK53a, whereas OsSWI3C does not interact with OsWAK53a. The bimolecular fluorescence complementation experiments show that OsSK4 and OsWAK50 and OsWAK53a can interact with each other in plants.The above results demonstrate that OsWAK50 The molecular mechanism of functional function provides important clues.