Protein-protein interactions and glycerophospholipids in bromovirus and nodavirus RNA replication.
Protein-protein interactions and glycerophospholipids in bromovirus and nodavirus RNA replication.
复制标题
溴病毒和诺达病毒 RNA 复制中的蛋白质-蛋白质相互作用和甘油磷脂。
DOI：
10.1007/978-3-7091-9326-6_14
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Hershberger,R
中科院分区：
文献类型：
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作者：
Ahlquist,P;Wu,SX;Kaesberg,P;Kao,CC;Quadt,R;DeJong,W;Hershberger,R
植物溴病毒和动物野田病毒是不同的正链RNA病毒，已被证明是RNA复制研究的有用模型。溴病毒编码RNA复制所需的两个大蛋白质：1a含有涉及解旋酶和加帽功能的结构域，2a含有中心聚合酶样结构域。使用免疫沉淀和远蛋白质印迹，我们现在已经表明，la和2a形成一个特定的复合物在体外，并绘制了相互作用的结构域。分子遗传学数据表明1a-2a复合体参与RNA复制，并表明它支持假定的解旋酶、聚合酶和加帽结构域的协调作用。相互作用的la和2a结构域的位置对复制模型和携带融合与分裂形式的同源复制基因的病毒基因组的进化具有影响。对于野田村病毒群舍病毒（FHV），已经分离出一种真正的RNA复制酶，其对添加的FHV RNA进行完全的、高活性的复制，以主要的ssRNA形式产生新合成的正链RNA。在该FHV无细胞系统中的正链RNA合成强烈依赖于添加几种甘油磷脂中的任何一种。正链RNA的合成取决于完整的甘油磷脂结构，包括极性头基和二酰基甘油脂质部分，并受到酰基链长度的强烈影响。
The plant bromoviruses and animal nodaviruses are distinct groups of positive strand RNA viruses that have proven to be useful models for RNA replication studies. Bromoviruses encode two large proteins required for RNA replication: 1a contains domains implicated in helicase and capping functions, and 2a contains a central polymerase-like domain. Using immunoprecipitation and far-western blotting, we have now shown that la and 2a form a specific complex in vitro and have mapped the interacting domains. Molecular genetic data implicate the 1a–2a complex in RNA replication and suggest that it supports coordinate action of the putative helicase, polymerase, and capping domains. The locations of the interacting la and 2a domains have implications for replication models and the evolution of virus genomes bearing homologous replication genes in fused vs. divided forms. For the nodavirus Flock house virus (FHV), a true RNA replicase has been isolated that carries out complete, highly active replication of added FHV RNA, producing newly synthesized positive strand RNA in pre-dominantly ssRNA form. Positive strand RNA synthesis in this FHV cell-free system is strongly dependent on the addition of any of several glycerophospholipids. Positive strand RNA synthesis depends on the complete glycerophospholipid structure, including the polar head group and diacyl glycerol lipid portion, and is strongly influenced by acyl chain length.