马腺苷酸脱氨酶1在马胎儿皮肤细胞中促进马传染性贫血病毒LTR的启动活性  

Over-expression of equine ADAR1 enhances the activity of equine infectious anemia virus LTR in FED cells

作　　者：付丽华[1] 汤艳东[2] 那雷[2] 王雪峰[2] 林跃智[2] 杜承[2] 周建华[2] 曲娟娟[1] 

机构地区：[1]东北农业大学生命科学学院,黑龙江哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/大动物病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2014年第1期11-14,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家十二五"艾滋病和病毒性肝炎等重大传染病防治"科技重大专项(2012ZX10001008-004);国家自然科学基金项目(31070809);国家自然科学青年基金项目(30901349)

摘　　要：腺苷酸脱氨酶(ADAR)1可催化双链RNA的腺嘌呤核苷酸(A)脱氨转化为次黄嘌呤核苷酸(I),该基因参与了很多病毒的复制与进化。在前期研究中对马传染性贫血病毒(EIAV)疫苗株基因组在体外培养细胞传代致弱过程中的变异规律进行了分析,表明与致弱前强毒株相比,EIAV弱毒疫苗株基因组出现高频率的A-I超突变,推测主要由腺苷酸脱氨酶1(ADAR1)催化导致,使基因组突变累积后的EIAV体内复制能力降低而弱化。为初步研究ADAR1在EIAV致弱中的作用,本研究应用PCR技术首次克隆了马ADAR1(eADAR1)的cDNA。经确认该重组蛋白在293T细胞中的正确表达后,将表达eADAR1的质粒与EIAV长末端重复区(LTR)控制的报告基因质粒共同转染马胎皮肤(FED)细胞,检测eADAR1对LTR启动活性的影响。实验数据表明,该ADAR1可以在体外培养的EIAV靶细胞中显著促进LTR启动的荧光素酶表达,显示eADAR1可以通过调控LTR促进EIAV的复制。以上结果提示eADAR1可以明显影响EIAV复制,但其机制较复杂,尚待深入研究。To investigate the equine infectiou anemia virus （EIAV） attenuated mechanism, we compared the genomes of virulent strains and vaccines, and the result showed that high frequency of A to G hypermutation was identified in viral genomes of EIAV during the attenuating process. Base on the published results of others, it is presumed that the adenosine deaminases that Act on RNA 1 （ADAR1） played an important role in the A to G hypermutation. And we speculated the accumulated A to G mutation might reduce the EIAV replication ability which attenuated the virus. In this study, to explore the role of ADAR1, the equine ADAR1 cDNA was cloned and over-expressed in cultivated equine fetal dermal （FED） cells. The results showed that the recombinant ADAR1 significantly enhanced the luciferase activity in a reporter plasmid that promoted by EIAV LTR, suggesting the enhancive potency rather than inhibition of equine ADAR1 on EIAV replication. These results suggest thateADAR1 was able to significantly affect EIAV replication, but the mechanism is more complex, have yet to be explored.

关 键 词：马腺苷酸脱氨酶1 马传染性贫血病毒 长末端重复区 

分 类 号：S852.65[农业科学—基础兽医学]