In vivo analysis of the TSWV cap-snatching mechanism:: single base complementarity and primer length requirements
In vivo analysis of the TSWV cap-snatching mechanism:: single base complementarity and primer length requirements
复制标题
DOI：
10.1093/emboj/20.10.2545
复制
发表时间：
2001-05-15
期刊：
影响因子：
11.4
通讯作者：
Goldbach, R
中科院分区：
文献类型：
--
作者：
Duijsings, D;Kormelink, R;Goldbach, R
在表达AMV复制酶蛋白的转基因烟草植物中，使用突变体苜蓿花叶病毒（AMV）RNA作为特异性帽供体，测试了在番茄斑萎病毒（TSWV）转录起始期间使用的加帽前导序列的要求。使用一系列在5 '非翻译区或亚基因组RNA4前导区中修饰的AMV RNA3突变体，序列分析显示切割的前导区长度可以在13和18个核苷酸之间变化。切割优先发生在A残基，表明需要与TSWV RNA模板的单碱基互补性，这可以通过分析体内用作帽供体的宿主mRNA来证实。
Requirements for capped leader sequences for use during transcription initiation by tomato spotted wilt virus (TSWV) were tested using mutant alfalfa mosaic virus (AMV) RNAs as specific cap donors in transgenic Nicotiana tabacum plants expressing the AMV replicase proteins. Using a series of AMV RNA3 mutants modified in either the 5'-non-translated region or in the subgenomic RNA4 leader, sequence analysis revealed that cleaved leader lengths could vary between 13 and 18 nucleotides. Cleavage occurred preferentially at an A residue, suggesting a requirement for a single base complementarity with the TSWV RNA template, which could be confirmed by analyses of host mRNAs used in vivo as cap donors.