Subgenomic RNA as a riboregulator: negative regulation of RNA replication by Barley yellow dwarf virus subgenomic RNA 2.
Subgenomic RNA as a riboregulator: negative regulation of RNA replication by Barley yellow dwarf virus subgenomic RNA 2.
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DOI：
10.1016/j.virol.2004.06.025
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发表时间：
2004-10
期刊：
影响因子：
3.7
通讯作者：
Ruizhong Shen;W. Miller
中科院分区：
文献类型：
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作者：
Ruizhong Shen;W. Miller
大麦黄矮病毒（Barley yellow dwarf virus，BYDV）在感染细胞中产生3个3′-coteminal subgenome RNA（sgRNA）。BYDV基因组RNA（gRNA）和sgRNA 1的翻译由3′端非翻译区的非帽依赖性翻译元件（BTE）介导。sgRNA 2和3不太可能是mRNA。我们认为，在其5′ UTR中含有BTE的sgRNA 2的积累通过反式抑制来自基因组RNA（gRNA）的病毒聚合酶的翻译来调节BYDV的复制。在此，我们测试了该假设并发现：（i）BTE或sgRNA 2与BYDV RNA的共接种抑制原生质体中的BYDV RNA积累;（ii）工程化以含有BTE的雀麦花叶病毒（BMV）反式抑制BYDV复制;和（iii）在BYDV感染期间产生的sgRNA 2反式抑制来自BMV RNA的GFP表达和非病毒报告mRNA的翻译。我们得出结论，sgRNA 2，通过其BTE，作为核糖核酸调节子抑制gRNA的翻译。这可以使gRNA可用作复制酶模板和用于双糖苷化。因此，BYDV sgRNA 2加入了越来越多的反式作用调节RNA的列表。
Barley yellow dwarf virus (BYDV) generates three 3′-coterminal subgenomic RNAs (sgRNAs) in infected cells. Translation of BYDV genomic RNA (gRNA) and sgRNA1 is mediated by the BYDV cap-independent translation element (BTE) in the 3′ untranslated region. sgRNAs 2 and 3 are unlikely to be mRNAs. We proposed that accumulation of sgRNA2, which contains the BTE in its 5′ UTR, regulates BYDV replication by trans-inhibiting translation of the viral polymerase from genomic RNA (gRNA). Here, we tested this hypothesis and found that: (i) co-inoculation of the BTE or sgRNA2 with BYDV RNA inhibits BYDV RNA accumulation in protoplasts; (ii) Brome mosaic virus (BMV), engineered to contain the BTE, trans-inhibits BYDV replication; and (iii) sgRNA2 generated during BYDV infection trans-inhibits both GFP expression from BMV RNA and translation of a non-viral reporter mRNA. We conclude that sgRNA2, via its BTE, functions as a riboregulator to inhibit translation of gRNA. This may make gRNA available as a replicase template and for encapsidation. Thus, BYDV sgRNA2 joins a growing list of trans-acting regulatory RNAs.