Authentic in vitro replication of two tombusviruses in isolated mitochondrial and endoplasmic reticulum membranes.
Authentic in vitro replication of two tombusviruses in isolated mitochondrial and endoplasmic reticulum membranes.
复制标题
两种烟草病毒在分离的线粒体和内质网膜中的真实体外复制。
DOI：
10.1128/jvi.00973-12
复制
发表时间：
2012
影响因子：
5.4
通讯作者：
Nagy,PeterD
中科院分区：
文献类型：
--
作者：
Xu,Kai;Huang,Tyng-Shyan;Nagy,PeterD
Replication of plus-stranded RNA viruses takes place on membranous structures derived from various organelles in infected cells. Previous works with Tomato bushy stunt tombusvirus (TBSV) revealed the recruitment of either peroxisomal or endoplasmic reticulum (ER) membranes for replication. In case of Carnation Italian ringspot tombusvirus (CIRV), the mitochondrial membranes supported CIRV replication. In this study, we developed ER and mitochondrion-basedin vitrotombusvirus replication assays. Using purified recombinant TBSV and CIRV replication proteins, we showed that TBSV could use the purified yeast ER and mitochondrial preparations for complete viral RNA replication, while CIRV preferentially replicated in the mitochondrial membranes. The viral RNA became partly RNase resistant after ∼40 to 60 min of incubation in the purified ER and mitochondrial preparations, suggesting that assembly of TBSV and CIRV replicases could take place in the purified ER and mitochondrial membranesin vitro. Using chimeric and heterologous combinations of replication proteins, we showed that multiple domains within the replication proteins are involved in determining the efficiency of tombusvirus replication in the two subcellular membranes. Altogether, we demonstrated that TBSV is less limited while CIRV is more restricted in utilizing various intracellular membranes for replication. Overall, the current work provides evidence that tombusvirus replication could occurin vitroin isolated subcellular membranes, suggesting that tombusviruses have the ability to utilize alternative organellar membranes during infection that could increase the chance of mixed virus replication and rapid evolution during coinfection.
