Cornell University, Plant Pathology and Plant Microbe-Biology Section, School of Integrative Plant Science, Cornell AgriTech at the New York State Agricultural Experiment Station, Geneva, NY 14456, USA.
Present address: Department of Biology, Utica College, Utica, NY 13502, USA.
J Gen Virol. 2021 May;102(5). doi: 10.1099/jgv.0.001607.
The RNA-dependent RNA polymerase (1E) is involved in replication of grapevine fanleaf virus (GFLV, , ) and causes vein clearing symptoms in . Information on protein 1E interaction with other viral and host proteins is scarce. To study protein 1E biology, three GFLV infectious clones, i.e. GHu (a symptomatic wild-type strain), GHu-1E (an asymptomatic GHu mutant) and F13 (an asymptomatic wild-type strain), were engineered with protein 1E fused to a V5 epitope tag at the C-terminus. Following -mediated delivery of GFLV clones in and protein extraction at seven dpi, when optimal 1E:V5 accumulation was detected, two viral and six plant putative interaction partners of V5-tagged protein 1E were identified for the three GFLV clones by affinity purification and tandem mass spectrometry. This study provides insights into the protein interactome of 1E during GFLV systemic infection in and lays the foundation for validation work.
RNA 依赖性 RNA 聚合酶（1E）参与葡萄扇叶病毒（GFLV，，）的复制，并导致感病的叶片出现脉带褪绿症状。关于 1E 蛋白与其他病毒和宿主蛋白相互作用的信息很少。为了研究 1E 蛋白的生物学功能，构建了三个 GFLV 感染性克隆，即 GHu（一种有症状的野生型毒株）、GHu-1E（一种无症状的 GHu 突变体）和 F13（一种无症状的野生型毒株），它们在 C 末端融合了 V5 表位标签。通过瞬时转染本氏烟并在 7dpi 提取蛋白，当检测到最佳的 1E:V5 积累时，利用亲和纯化和串联质谱法鉴定了三个 GFLV 克隆中 V5 标记的 1E 蛋白的两个病毒和六个宿主潜在互作伙伴。本研究为 GFLV 在本氏烟中的系统感染过程中 1E 的蛋白互作组提供了新的见解，并为后续的验证工作奠定了基础。
