VP16 INTERACTS VIA ITS ACTIVATION DOMAIN WITH VP22, A TEGUMENT PROTEIN OF HERPES-SIMPLEX VIRUS, AND IS RELOCATED TO A NOVEL MACROMOLECULAR ASSEMBLY IN COEXPRESSING CELLS
VP16 INTERACTS VIA ITS ACTIVATION DOMAIN WITH VP22, A TEGUMENT PROTEIN OF HERPES-SIMPLEX VIRUS, AND IS RELOCATED TO A NOVEL MACROMOLECULAR ASSEMBLY IN COEXPRESSING CELLS
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DOI：
10.1128/jvi.69.12.7932-7941.1995
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发表时间：
1995-12-01
影响因子：
5.4
通讯作者：
OHARE, P
中科院分区：
文献类型：
--
作者：
ELLIOTT, G;MOUZAKITIS, G;OHARE, P
除了作为病毒立即早期转录的强大反式激活因子的功能外，VP16 也是单纯疱疹病毒 (HSV) 病毒粒子的重要组成部分。因此，VP16 作为病毒外皮的一部分被引入细胞中，以实现其反式激活功能。在这里，我们检查了病毒感染细胞特异的 VP16 蛋白质-蛋白质相互作用的潜力，并表明 VP16 在高度富集的级分中与单一主要多肽（我们将其鉴定为病毒编码的结构蛋白 VP22）共纯化。我们进一步表明，体外翻译的 VP22 与纯化的 VP16 特异性结合。 VP16 的激活结构域对于这种结合是必需的并且基本上足够了。该结构域内的突变会破坏其反式激活功能，也会影响 VP22 的结合。此外，我们发现，虽然 VP16 和 VP22 在体内表现出不同的区室化模式，但这两种蛋白质的共表达导致了从正常位置到新型大分子组装的深刻重组。共定位还依赖于 VP16 的激活结构域，但需要 N 末端内的额外决定簇。这些结果在 VP16 转录调节的背景下进行了讨论，包括感染早期皮膜蛋白递送期间和病毒组装后期。
In addition to its function as a powerful transactivator of viral immediate-early transcription, VP16 is an essential component of the herpes simplex virus (HSV) virion. As such, VP16 is introduced into cells, to effect its function in transactivation, as part of the virus tegument. Here we examine the potential for VP16 protein-protein interactions specific to virus-infected cells and show that VP16 copurifies in a highly enriched fraction with a single major polypeptide which we identify as the virus-encoded structural protein VP22. We further show that in vitro-translated VP22 binds specifically to purified VP16. The activation domain of VP16 was required and largely sufficient for this binding. Mutations within this domain, which disrupt its transactivation function, also affected VP22 binding. Furthermore, we show that while VP16 and VP22 showed distinct patterns of compartmentalization in vivo, coexpression of both proteins resulted in a profound reorganization from their normal locations to a novel macromolecular assembly. The colocalization was also dependent on the activation domain of VP16 but required additional determinants within the N terminus. These results are discussed in the context of VP16 regulation of transcription both early in infection during delivery of tegument proteins and at late times during virus assembly.