Binding of tobamovirus replication protein with small RNA duplexes
Binding of tobamovirus replication protein with small RNA duplexes
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DOI：
10.1099/vir.0.82994-0
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发表时间：
2007-08-01
影响因子：
3.8
通讯作者：
Watanabe, Yuichiro
中科院分区：
文献类型：
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作者：
Kurihara, Yukio;Inaba, Naoko;Watanabe, Yuichiro
用小RNA克隆技术测定了感染烟草花叶病毒(TMV)-CG的拟南芥的小干扰RNA(SiRNAs)序列。大多数TIMV来源的siRNAs的长度为21nT。最丰富的内源小RNA大小在TMV侵染植株中为21nt，而在模拟接种植株中为24nt。Northern印迹分析表明，一些microRNAs(MiRNAs)在TMV侵染植株中比在模拟接种植株中积累得更多。研究了TMV-CG编码的126K复制蛋白(一种RNA沉默抑制因子)是否导致小RNA浓缩的问题。复制蛋白的瞬时表达并没有改变miRNA的加工模式。然而，miRNA、miRNA*(miRNA双链的相反链)和发夹来源的siRNA都与复制蛋白共沉淀。凝胶迁移率-位移分析表明，该复制蛋白结合了小的RNA双链。这些结果表明，Tabamovinus复制蛋白通过结合小RNA双链，改变受感染植物的小RNA图谱，发挥沉默抑制的作用。
The sequence profiles of small interfering RNAs (siRNAs) in Arabidopsis infected with the crucifer tobamovirus tobacco mosaic virus (TMV)-Cg were determined by using a small RNA cloning technique. The majority of TIMV-derived siRNAs were 21 nt in length. The size of the most abundant endogenous small RNAs in TMV-infected plants was 21 nt, whilst in mock-inoculated plants, it was 24 nt. Northern blot analysis revealed that some microRNAs (miRNAs) accumulated more in TMV-infected plants than in mock-inoculated plants. The question of whether the TMV-Cg-encoded 126K replication protein, an RNA-silencing suppressor, caused small RNA enrichment was examined. Transient expression of the replication protein did not change the pattern of miRNA processing. However, miRNA, miRNA* (the opposite strand of the miRNA duplex) and hairpin-derived siRNA all co-immunoprecipitated with the replication protein. Gel mobility-shift assays indicated that the replication protein binds small RNA duplexes. These results suggest that the tobamovinus replication protein functions as a silencing suppressor by binding small RNA duplexes, changing the small RNA profile in infected plants.