Hofer J M, Dekker E L, Reynolds H V, Woolston C J, Cox B S, Mullineaux P M
John Innes Institute, John Innes Centre for Plant Science Research, Norwich, United Kingdom.
Plant Cell. 1992 Feb;4(2):213-23. doi: 10.1105/tpc.4.2.213.
We have investigated the relationship between viral DNA replication and virion sense gene expression in wheat dwarf virus (WDV), a member of the geminivirus group, by testing a series of deletion mutants in transfected Triticum monococcum (einkorn) protoplasts. Mutants contained a transcription fusion of the chloramphenicol acetyltransferase coding sequence to the virion sense promoter that replaced the viral coat protein coding sequence. The deletion analysis revealed that WDV replication and virion sense transcription can proceed independently and are controlled in part by nonoverlapping elements in the large intergenic region. These data and those from a C2 open reading frame (ORF) frameshift mutant also showed that the product of the C2 ORF (C1-C2 protein) is independently involved in both DNA replication and activation of the virion sense promoter. The amino acid sequences encoded by C2, which are highly conserved in the geminivirus group, show some homology to the DNA binding domain of the myb-related class of plant transcription factors. The possible involvement of the host in controlling the function of the C1-C2 protein and the implication of these data for the development of WDV-based gene vectors are discussed.
我们通过在转染的一粒小麦原生质体中测试一系列缺失突变体,研究了双生病毒组成员小麦矮缩病毒(WDV)中病毒DNA复制与病毒链基因表达之间的关系。突变体包含氯霉素乙酰转移酶编码序列与病毒链启动子的转录融合体,该启动子取代了病毒外壳蛋白编码序列。缺失分析表明,WDV复制和病毒链转录可以独立进行,并且部分受大基因间隔区中不重叠元件的控制。这些数据以及来自C2开放阅读框(ORF)移码突变体的数据还表明,C2 ORF的产物(C1-C2蛋白)独立参与DNA复制和病毒链启动子的激活。C2编码的氨基酸序列在双生病毒组中高度保守,与植物转录因子中myb相关类别的DNA结合域有一些同源性。讨论了宿主可能参与控制C1-C2蛋白的功能以及这些数据对基于WDV的基因载体开发的意义。