A nuclear localization of the infectious haematopoietic necrosis virus NV protein is necessary for optimal viral growth.
A nuclear localization of the infectious haematopoietic necrosis virus NV protein is necessary for optimal viral growth.
复制标题
DOI：
10.1371/journal.pone.0022362
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Park JW
中科院分区：
文献类型：
--
作者：
Choi MK;Moon CH;Ko MS;Lee UH;Cho WJ;Cha SJ;Do JW;Heo GJ;Jeong SG;Hahm YS;Harmache A;Bremont M;Kurath G;Park JW
先前报道，传染性造血组织坏死病毒 (IHNV) 的非病毒体 (NV) 蛋白对于 IHNV 的有效生长和致病性至关重要。然而，人们对 NV 支持病毒生长的机制知之甚少。在本研究中，研究了 NV 的细胞定位及其在宿主细胞中 IHNV 生长中的作用。通过瞬时转染与绿色荧光蛋白 (GFP) 融合的 NV 的 RTG-2 细胞，证明了 NV 的核定位。缺失分析表明，32EGDL35 残基对于 NV 蛋白的核定位至关重要，这 4 个氨基酸与 GFP 的融合引导其转运至细胞核。我们生成了重组 IHNV，rIHNV-NV-ΔEGDL，其中 32EGDL35 从 NV 中删除。具有野生型 NV (rIHNV-NV) 或 NV 基因替换为 GFP (rIHNV-ΔNV-GFP) 的 rIHNV 用作对照。在感染后 48 小时 (p.i.)，用 rIHNV-ΔNV-GFP 和 rIHNV-NV-ΔEGDL 感染的 RTG-2 细胞产生的感染性病毒颗粒分别比野生型 rIHNV 感染细胞少 12 倍和 5 倍。注射后 24 小时用 Poly I∶C 处理。 Poly I∶C 不抑制野生型 rIHNV 的复制，但 rIHNV-ΔNV-GFP 和 rIHNV-NV-ΔEGDL 的复制速率受到抑制。此外，rIHNV-ΔNV和rIHNV-NV-ΔEGDL均诱导比野生型rIHNV更高水平的IFN1和Mx1表达。这些数据表明，IHNV NV可能通过抑制INF系统来支持IHNV的生长，并且负责核定位的32EGDL35的氨基酸残基对于NV的抑制活性很重要。
The nonvirion (NV) protein of infectious hematopoietic necrosis virus (IHNV) has been previously reported to be essential for efficient growth and pathogenicity of IHNV. However, little is known about the mechanism by which the NV supports the viral growth. In this study, cellular localization of NV and its role in IHNV growth in host cells was investigated. Through transient transfection in RTG-2 cells of NV fused to green fluorescent protein (GFP), a nuclear localization of NV was demonstrated. Deletion analyses showed that the 32EGDL35 residues were essential for nuclear localization of NV protein, and fusion of these 4 amino acids to GFP directed its transport to the nucleus. We generated a recombinant IHNV, rIHNV-NV-ΔEGDL in which the 32EGDL35 was deleted from the NV. rIHNVs with wild-type NV (rIHNV-NV) or with the NV gene replaced with GFP (rIHNV-ΔNV-GFP) were used as controls. RTG-2 cells infected with rIHNV-ΔNV-GFP and rIHNV-NV-ΔEGDL yielded 12- and 5-fold less infectious virion, respectively, than wild type rIHNV-infected cells at 48 h post-infection (p.i.). While treatment with poly I∶C at 24 h p.i. did not inhibit replication of wild-type rIHNVs, replication rates of rIHNV-ΔNV-GFP and rIHNV-NV-ΔEGDL were inhibited by poly I∶C. In addition, both rIHNV-ΔNV and rIHNV-NV-ΔEGDL induced higher levels of expressions of both IFN1 and Mx1 than wild-type rIHNV. These data suggest that the IHNV NV may support the growth of IHNV through inhibition of the INF system and the amino acid residues of 32EGDL35 responsible for nuclear localization are important for the inhibitory activity of NV.
登录
查看更多内容
影响因子：
5
作者：
Hiscox JA
通讯作者：
Hiscox JA
影响因子：
3.7
作者：
Haller O;Kochs G;Weber F
通讯作者：
Weber F
影响因子：
56.9
作者：
Enninga, J;Levy, DE;Fontoura, BMA
通讯作者：
Fontoura, BMA
影响因子：
5.4
作者：
Brzózka, K;Finke, S;Conzelmann, KK
通讯作者：
Conzelmann, KK
影响因子：
5.4
作者：
KURATH, G;AHERN, KG;LEONG, JC
通讯作者：
LEONG, JC