Centex Shrimp, Faculty of Science, Mahidol University, Bangkok, Thailand.
PLoS One. 2011;6(9):e25420. doi: 10.1371/journal.pone.0025420. Epub 2011 Sep 29.
White spot syndrome virus (WSSV) is one of the most serious pathogens of penaeid shrimp. Although its genome has been completely characterized, the functions of most of its putative proteins are not yet known. It has been suggested that the major nucleocapsid protein VP15 is involved in packaging of the WSSV genome during virion formation. However, little is known in its relationship with shrimp host cells. Using the yeast two-hybrid approach to screen a shrimp lymphoid organ (LO) cDNA library for proteins that might interact with VP15, a protein named PmFKBP46 was identified. It had high sequence similarity to a 46 kDa-immunophilin called FKBP46 from the lepidopteran Spodoptera frugiperda (the fall armyworm). The full length PmFKBP46 consisted of a 1,257-nucleotide open reading frame with a deduced amino acid sequence of 418 residues containing a putative FKBP-PPIase domain in the C-terminal region. Results from a GST pull-down assay and histological co-localization revealed that VP15 physically interacted with PmFKBP46 and that both proteins shared the same subcellular location in the nucleus. An electrophoretic mobility shift assay indicated that PmFKBP46 possessed DNA-binding activity and functionally co-interacted with VP15 in DNA binding. The overall results suggested that host PmFKBP46 might be involved in genome packaging by viral VP15 during virion assembly.
白斑综合征病毒(WSSV)是对虾最重要的病原体之一。尽管其基因组已被完全表征,但大多数假定蛋白的功能仍未知。有人提出,主要核衣壳蛋白 VP15 参与了病毒粒子形成过程中 WSSV 基因组的包装。然而,其与虾宿主细胞的关系知之甚少。本研究利用酵母双杂交方法筛选虾淋巴器官(LO)cDNA 文库中可能与 VP15 相互作用的蛋白,鉴定到一个名为 PmFKBP46 的蛋白。它与鳞翅目昆虫草地贪夜蛾(the fall armyworm)中的 46 kDa 免疫亲和素 FKBP46 具有高度的序列相似性。全长 PmFKBP46 由一个 1257 个核苷酸的开放阅读框组成,推测其编码 418 个氨基酸,在 C 末端区域含有一个推定的 FKBP-PPIase 结构域。GST 下拉测定和组织学共定位结果表明,VP15 与 PmFKBP46 发生物理相互作用,并且这两种蛋白在细胞核中具有相同的亚细胞定位。电泳迁移率变动分析表明,PmFKBP46 具有 DNA 结合活性,并与 VP15 在 DNA 结合中具有功能上的相互作用。总体结果表明,宿主 PmFKBP46 可能参与了病毒 VP15 在病毒粒子组装过程中对基因组的包装。