两个高度同源的RING结构域蛋白功能初步研究  

Initial Studies on Protein Function of Two Highly Homologous RING Structure

作　　者：杨虹伟[1] 翟先芝 刘志斌[1] 王健美[1] 李旭锋[1] 杨毅[1] 

机构地区：[1]四川大学生命科学学院,生物资源与生态环境教育部重点实验室,成都610064

出　　处：《中国农业科技导报》2013年第5期100-105,共6页Journal of Agricultural Science and Technology

基　　金：国家自然科学基金项目(31171586);国家转基因生物新品种培育重大专项(2011ZX08009-003-002);中央高校基本科研业务费项目(2011SCU11107)资助

摘　　要：泛素化途径在植物应答生物和非生物胁迫中起着重要作用,其中E3连接酶在泛素化途径中起着决定性作用。室通过在线基因芯片预测发现2个与非生物胁迫相关的拟南芥RING结构域蛋白基因At2g24480(HHR1)、At5g43200(HHR2)通过体外泛素化实验证明了二者具有E3连接酶活性。对这两个E3连接酶进行了亚细胞定位,表达谱和组织特异性分析,发现二者受诱导后的表达情况、组织特异性、亚细胞定位均不同。HHR1定位于细胞膜上,HHR2定位于细胞核;表达谱分析表明HHR1响应热和H2O2胁迫,而HHR2响应盐和H2O2胁迫;HHR1在根和花中的表达量较高,在茎和叶中的表达量较低,而HHR2在根、茎、叶中的表达量都较高,在花中表达量较低。该研究结果证实了HHR1和HHR2的一些性质和亚细胞定位,为深入研究其功能奠定了基础。Ubiquitination pathway plays important roles in plant responses to biotic and abiotic stress, especially E3 plays decisive role in pathway of ubiquifin ligascs. 2 Arabidopsis RING domain protein At2g24480 （ HHR1 ） and At5943200（HHR2） were found, which were related to abiotic stress, through prediction of gene chip online. In vitro self-ubiquitination assay demonstrated that the 2 RING domain proteins possess an E3 ligase activity. In this paper, sub-cellular localization, gene expression profiles, tissue specificity of them were conducted. We found that they showed differences in sub-cellular localization, gene expression profiles, tissue specificity. HHR1 was positioned on cell membrane, while HHR2 in nucleus. Our studies demonstrated that both HHR1 and HHR2 were related to H202 stress. In addition, HHR1 and HHR2 were related to heat and salt stress, respectively. The analysis of expressional pattern revealed that HHR1 was highly expressed in root and flower, less expressed in leaves and stems. However, HHR2 relatively expressed less in flower, but highly expressed in roots, stems and leaves. These results confirmed that certain properties and sub-cellular localization of HHR1 and HHR2, and laid foundation for further studies on their function.

关 键 词：E3连接酶 亚细胞定位 表达谱分析 组织特异性 

分 类 号：Q786[生物学—分子生物学]