Replicative intermediates of Tomato leaf curl virus and its satellite DNAs
Replicative intermediates of Tomato leaf curl virus and its satellite DNAs
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DOI：
10.1016/j.virol.2004.10.043
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发表时间：
2005-01-20
期刊：
影响因子：
3.7
通讯作者：
Jeske, H
中科院分区：
文献类型：
--
作者：
Alberter, B;Rezaian, MA;Jeske, H
近年来，一些植物双生病毒已被证明利用滚环复制（RCR）和重组依赖性复制（RDR）策略。病毒复制相关蛋白（Rep）与其同源DNA的高度特异性结合对于启动病毒DNA复制和识别双生病毒属的双链双生病毒的DNA组分是必不可少的。我们已经扩大了复制分析的单部澳大利亚番茄曲叶病毒（ToLCV），其Rep结合缺陷突变体，卫星DNA支持。通过单链（SS）DNA选择性纤维素色谱分离后，通过二维琼脂糖凝胶电泳的病毒DNA的分析表明，ToLCV及其突变体的DNA中间体是相同的。鉴定了RCR和RDR中间体。观察到新的ToLCV DNA形式，并将其表征为亚基因组拓扑异构体、异质性开环双链（ds）DNA和降解产物。一个1350-nt的DNA β卫星与无关的棉花曲叶木尔坦病毒（CLCuMV）的支持ToLCV和产生的中间体的RCR和RDR，表明卫星的复制策略是由辅助病毒。682 nt的ToLCV卫星DNA的复制中间体不能被解决，然而，高达八聚体的多联体被检测到，连同一个领域的杂交材料暗示互补链复制的异质性环状ssDNA模板。(C)2004年爱思唯尔公司All rights reserved.
Several plant geminiviruses have been shown recently to utilize both rolling-circle replication (RCR) and recombination-dependent replication (RDR) strategies. A highly specific binding of the viral replication-associated protein (Rep) to its cognate DNA is essential for initiation of viral DNA replication and for the recognition of DNA components of the bipartite geminiviruses of the Begomovirus genus. We have extended the replication analysis to the monopartite Australian Tomato leaf curl virus (ToLCV), its Rep binding deficient mutants, and the satellite DNAs it supports. Analyses of viral DNA by two-dimensional agarose gel electrophoresis after fractionation by single-stranded (ss) DNA-selective cellulose chromatography revealed that DNA intermediates of ToLCV and its mutant were identical. Both RCR and RDR intermediates were identified. New ToLCV DNA forms were observed and characterized as subgenomic topoisomers, heterogeneous open circular double-stranded (ds) DNA, and degradation products. A 1350-nt DNA beta satellite associated with the unrelated Cotton leaf curl Multan virus (CLCuMV) was supported by ToLCV and produced intermediates of both RCR and RDR, suggesting that replication strategies of satellites are determined by the helper virus. Replicative intermediates of the 682 nt ToLCV satellite DNA could not be resolved; however, concatemers of up to octamer were detected, together with a field of hybridizing material suggestive of complementary strand replication on heterogeneous circular ssDNA templates. (C) 2004 Elsevier Inc. All rights reserved.