Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada.
J Virol. 2011 Oct;85(19):9696-707. doi: 10.1128/JVI.00833-11. Epub 2011 Jul 20.
me53 is an immediate-early/late gene found in all lepidopteran baculoviruses sequenced to date. Deletion of me53 results in a greater-than-1,000-fold reduction in budded-virus production in tissue culture (J. de Jong, B. M. Arif, D. A. Theilmann, and P. J. Krell, J. Virol. 83:7440-7448, 2009). We investigated the localization of ME53 using an ME53 construct fused to green fluorescent protein (GFP). ME53:GFP adopted a primarily cytoplasmic distribution at early times postinfection and a primarily nuclear distribution at late times postinfection. Additionally, at late times ME53:GFP formed distinct foci at the cell periphery. These foci colocalized with the major envelope fusion protein GP64 and frequently with VP39 capsid protein, suggesting that these cell membrane regions may represent viral budding sites. Deletion of vp39 did not influence the distribution of ME53:GFP; however, deletion of gp64 abolished ME53:GFP foci at the cell periphery, implying an association between ME53 and GP64. Despite the association of ME53 and GP64, ME53 fractionated with the nucleocapsid only after budded-virus fractionation. Together these findings suggest that ME53 may be providing a scaffold that bridges the viral envelope and nucleocapsid.
me53 是一种在迄今测序的所有鳞翅目杆状病毒中都存在的早期/晚期基因。缺失 me53 会导致组织培养中的出芽病毒产量降低 1000 倍以上(J. de Jong、B. M. Arif、D. A. Theilmann 和 P. J. Krell,J. Virol. 83:7440-7448, 2009)。我们使用与绿色荧光蛋白 (GFP) 融合的 ME53 构建体来研究 ME53 的定位。ME53:GFP 在感染后早期呈主要细胞质分布,在感染后晚期呈主要核分布。此外,在晚期 ME53:GFP 在细胞边缘形成明显的焦点。这些焦点与主要包膜融合蛋白 GP64 共定位,并经常与 VP39 衣壳蛋白共定位,表明这些细胞膜区域可能代表病毒出芽部位。VP39 的缺失不影响 ME53:GFP 的分布;然而,GP64 的缺失消除了细胞边缘的 ME53:GFP 焦点,暗示 ME53 与 GP64 之间存在关联。尽管 ME53 与 GP64 相关联,但 ME53 仅在出芽病毒分离后才与核衣壳分离。这些发现表明,ME53 可能提供了一种支架,将病毒包膜和核衣壳连接起来。