拟南芥BDR6的生物信息学分析及调控种子寿命的互作蛋白鉴定  

Bioinformatics analysis and interaction protein identification of BDR6,regulating seed longevity in Arabidopsis

作　　者：吴唐娟 朱静静 王源秀[1] 马锐奇 陈灵 刘阳 刘华彬 王程亮 WU Tangjuan;ZHU Jingjing;WANG Yuanxiu;MA Ruiqi;CHEN Ling;LIU Yang;LIU Huabin;WANG Chengliang(College of Life Sciences,Anhui Normal University,Wuhu,Anhui 241002,China;State Laboratory of Crop Stress Adaption and Improvement,Henan University,Kaifeng,Henan 475004,China;College of Life and Health Sciences,Anhui Science and Technology University,Bengbu,Anhui 233030,China;The Institute for Advanced Studies,Wuhan University,Wuhan 430072,China)

机构地区：[1]安徽师范大学生命科学学院,安徽芜湖241002 [2]河南大学作物逆境适应与改良国家重点实验室,河南开封475004 [3]安徽科技学院生命与健康科学学院,安徽蚌埠233130 [4]武汉大学高等研究院,武汉430072

出　　处：《植物生理学报》2024年第10期1548-1560,共13页Plant Physiology Journal

基　　金：国家自然科学基金(32101646);安徽省科学技术厅重点研发项目(2023n06020004);中国博士后科学基金项目(348510);芜湖市科学技术局应用基础研究项目(2022jc10)。

摘　　要：未知功能域(DUF,domain of unknown function)是一种未被表征功能的蛋白质结构域,DUFs基因在植物的生长、发育、繁殖和物质代谢进程中发挥着至关重要的作用。在拟南芥中有9个DUF620同源基因(BDR1~BDR9),该研究利用生物信息学、人工加速老化、亚细胞定位及酵母双杂交等技术对拟南芥BDR6基因进行了功能分析与研究。结果显示:拟南芥9个BDR蛋白的基序分布十分相近,具有相似的结构域;系统进化分析发现BDR6与杨树基因Potri.010G195200和Potri.008G062500亲缘关系较近,与BDR1处于同一大类分支;氨基酸结构预测发现BDR6由α-螺旋、延伸链和不规则卷曲组成,是不稳定的非跨膜的具有内在无序区域的亲水蛋白,定位于细胞质和细胞核中,在拟南芥种子发育的中后期显著表达;BDR6基因不具有转录因子结构域,启动子序列上具有特异的光响应元件G-box等多种顺式作用元件。人工加速老化结果显示,bdr6突变体种子发芽率显著高于野生型,说明BDR6负调控种子寿命。种子GUS染色和亚细胞定位结果均与生信预测结果一致。利用酵母双杂交技术筛选到5个与BDR6互作的蛋白,并进行一对一验证,结果发现EML3与BDR6蛋白有较强的互作。以上结果为研究BDR6基因的功能及其在拟南芥种子寿命中的调控作用提供了重要的理论基础。DUF(Domain of Unknown Function)is a protein structural domain that was not characterized its function,and DUFs genes play a crucial role in plant growth,development,reproduction,and material metabolism processes.There are nine DUF620 genes(BDR1–BDR9)in Arabidopsis thaliana.In this study,bioinformatics,artificial accelerated aging,subcellular localization and yeast two-hybrid technology were used to predict and study the function of BDR6.Nine BDRs contained very similar motifs,with conserved domains.Phylogenetic analysis revealed that BDR6 was closely related to the Populus trichocarpa genes Potri.010G195200 and Potri.008G062500,and the same large group as BDR1.BDR6 protein was composed ofα-helix,extended chain and irregular coil,and was an unstable non-transmembrane hydrophilic protein with intrinsically disordered region.The subcellular localization was in the cytoplasm and nucleus,and could be expressed in the middle and late stages of seeds development of Arabidopsis.The BDR6 gene does not have a transcription factor domain,and the promoter sequence contained a variety of cis-acting elements such as specific light response element G-box.After aging,the germination rate of bdr6 seeds was significantly higher than that of the wild type,indicating negative regulation of seed longevity.The results of GUS staining and subcellular localization were consistent with the bioinformatics prediction.Five proteins interacting with BDR6 were screened using yeast two hybrid technology,and one-on-one validation was performed,revealing that EML3 interacted with BDR6.These findings provide a theoretical basis for studying the gene function of BDR6 and its regulatory role in Arabidopsis seed longevity.

关 键 词：DUF620 BDR6 生物信息学 种子寿命 酵母双杂交 蛋白互作 

分 类 号：Q943.2[生物学—植物学]