Effect of actinomycin D on replication of satellite tobacco ringspot virus RNA in plant protoplasts.
Effect of actinomycin D on replication of satellite tobacco ringspot virus RNA in plant protoplasts.
复制标题
放线菌素 D 对植物原生质体中卫星烟草环斑病毒 RNA 复制的影响。
DOI：
10.1016/0042-6822(90)90483-8
复制
发表时间：
1990
期刊：
影响因子：
3.7
通讯作者：
Bruening,G
中科院分区：
文献类型：
--
作者：
Buckley,B;Bruening,G
我们建立了一个由寄主、烟草环斑病毒（TobRV）和卫星烟草环斑病毒RNA（sTobRV RNA）组成的三组分系统，用于研究寄主组分或TobRV基因产物对卫星RNA繁殖周期的特异性贡献。通过电穿孔法将sTobRV和TobRV基因组RNA分别接种豇豆（Vigna unguiculata）原生质体。通过印迹杂交和将[14 C]尿苷掺入到具有sTobRV RNA电泳迁移率的材料中，检测到sTobRV RNA的增加。50 μg/ml放线菌素D（ActD）能有效地抑制未接种原生质体的RNA合成。在与sTobRV RNA和TobRV基因组RNA共接种后24或48小时，向原生质体中添加Act D对sTobRV RNA的积累几乎没有影响，而在共接种前24小时添加Act D可防止任何检测到的任一极性的sTobRV RNA的积累。我们的研究结果和以前的研究结果的RNA互补的卫星RNA在感染组织提取物表明，RNA依赖的RNA聚合酶是负责合成的sTobRV RNA。当早期加入时，Act D的强烈抑制作用意味着宿主因子在sTobRV RNA复制的早期阶段中的作用。
We have developed a three-component system of host, tobacco ringspot virus (TobRV), and satellite tobacco ringspot virus RNA (sTobRV RNA) for investigating the specific contributions of host components or TobRV gene products to the propagative cycle of satellite RNA. Cowpea (Vigna unguiculata) protoplasts were inoculated with sTobRV and TobRV genomic RNAs by electroporation. An increase in sTobRV RNA was detected both by blot hybridization and by incorporation of [14C]uridine into material with the electrophoretic mobility of sTobRV RNA. DNA-dependent RNA synthesis in uninoculated protoplasts was effectively inhibited by 50 μg/ml actinomycin D (Act D) in the medium. Addition of Act D to protoplasts 24 or 48 hr after coinoculation with sTobRV RNA and TobRV genomic RNAs had little effect on accumulation of sTobRV RNA, whereas addition at 24 hr prior to coinoculation prevented any detected accumulation of sTobRV RNA of either polarity. Our results and previous findings of RNA complementary to encapsidated satellite RNA in extracts of infected tissue suggest that an RNA-dependent RNA polymerase is responsible for the synthesis of sTobRV RNA. The strongly inhibitory effect of Act D when added early implies a role for a host factor in the early phase of sTobRV RNA replication.