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HC-Pro 3个参与抑制RNA沉默氨基酸位点的鉴定

Identificaion of three amino acid sites from Potato virus A HC-Pro invovlved in suppression of RNA silencing
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摘要 马铃薯Y病毒属病毒编码的辅助成分-蛋白酶(helper component-proteinase,HC-Pro)是第一个被鉴定的RNA沉默抑制因子。本研究通过定点突变的方法获得了马铃薯A病毒(Potato virus A,PVA)HC-Pro的3个突变体,利用农杆菌共浸润的方法分析了这些突变对HC-Pro抑制RNA沉默活性的影响。与野生型HC-Pro处理相比,Phe6、Asn11缺失突变体的处理中绿色荧光减弱,而Ile250-Gly251-Asn252(IGN)基序中的Ile和Gly分别突变为Asp和Glu的处理中观察不到绿色荧光。该结果表明Phe6、Asn11和IGN2503个位点均参与调控HC-Pro的抑制RNA沉默活性。 The helper component-proteinase （HC-Pro） encoded by potyviruses is the first identified RNA silencing suppressor. In the study, three mutants of Potato virus A （PVA） HC-Pro were obtained via site-directed mutagenesis and analyzed for their activities on RNA silencing via agroinfiltration. Compared with that of wild type HC-Pro, the treatment with deletion mutants of amino acids Phe6 and Asnn resulted in reduced green fluorescence, while treatment with substitution mutant of Ile250-Gly251-Asn252 motif to Asp250Glu251Asn252 resulted in no fluorescence. These results suggested that amino acids Phe6, Asnll and motif Ile250-Gly251-Asn252 are involved in modulating the RNA silencing suppression activity of HC-Pro.
作者 王洁 刘金亮 阴筱 高瑞 刘红梅 李向东 刘焕庭
机构地区 山东农业大学植物保护学院 吉林大学植物科学学院 山东农业大学生命科学学院 Scottish Structural Proteomics Facility(SSPF)
出处 《植物病理学报》 CAS CSCD 北大核心 2012年第3期267-273,共7页 Acta Phytopathologica Sinica
基金 国家自然科学基金资助项目(30971895 31011130031) 新世纪优秀人才支持计划(NCET-07-0520) 教育部博士学科点基金(200804340006)
关键词 HC-PRO 定点突变 抑制RNA沉默 农杆菌共浸润 HC-Pro site-directed mutagenesis RNA silencing suppression agroinfiltration
分类号 S432.41 [农业科学—植物病理学] Q78 [生物学—分子生物学]
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参考文献24

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植物病理学报
2012年 第3期

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