Heat shock 70 protein interaction with Turnip mosaic virus RNA-dependent RNA polymerase within virus-induced membrane vesicles
Heat shock 70 protein interaction with Turnip mosaic virus RNA-dependent RNA polymerase within virus-induced membrane vesicles
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DOI：
10.1016/j.virol.2007.12.014
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发表时间：
2008-04-25
期刊：
影响因子：
3.7
通讯作者：
Fortin, Marc G.
中科院分区：
文献类型：
--
作者：
Dufresne, Philippe J.;Thivierge, Karine;Fortin, Marc G.
采用串联亲和纯化方法对拟南芥中芜菁花叶病毒（TuMV） RNA依赖性RNA聚合酶（RdRp）细胞相互作用物进行鉴定。热休克同源蛋白70-3 （Hsc70-3）和聚(A)结合（PABP）宿主蛋白在体外被发现与RdRp相互作用。如前所述，Hsc70-3被重新分配到受感染植物的核和膜组分中，两种RdRp相互作用物使用RdRp特异性抗体从富含膜的提取物中共免疫沉淀。荧光标记的RdRp和Hsc70-3在烟叶中单独或联合表达时定位于细胞质和细胞核。然而，当它们与TuNW的膜结合蛋白6K-VPg-Pro共表达时，它们被重新分配到大的核周er来源的囊泡中。Hsc70-3与RdRp的结合可能发生在膜源性复制复合体中。因此，Hsc70-3和PABP2是复制酶复合体的潜在组成部分，可能在多病毒RdRp功能的调控中发挥重要作用。(c) 2007爱思唯尔公司版权所有。
Tandem affinity purification was used in Arabidopsis thaliana to identify cellular interactors of Turnip mosaic virus (TuMV) RNA-dependent RNA polymerase (RdRp). The heat shock cognate 70-3 (Hsc70-3) and poly(A)-binding (PABP) host proteins were recovered and shown to interact with the RdRp in vitro. As previously shown for PABP, Hsc70-3 was redistributed to nuclear and membranous fractions in infected plants and both RdRp interactors were co-immunoprecipitated from a membrane-enriched extract using RdRp-specific antibodies. Fluorescently tagged RdRp and Hsc70-3 localized to the cytoplasm and the nucleus when expressed alone or in combination in Nicotiana benthamiana. However, they were redistributed to large perinuclear ER-derived vesicles when co-expressed with the membrane binding 6K-VPg-Pro protein of TuNW. The association of Hsc70-3 with the RdRp could possibly take place in membrane-derived replication complexes. Thus, Hsc70-3 and PABP2 are potentially integral components of the replicase complex and could have important roles to play in the regulation of potyviral RdRp functions. (c) 2007 Elsevier Inc. All rights reserved.