Assessment of the subcellular localization of the herpes simplex virus structural protein VP22 in the absence of other viral gene products
Assessment of the subcellular localization of the herpes simplex virus structural protein VP22 in the absence of other viral gene products
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DOI：
10.1016/s0168-1702(01)00355-0
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发表时间：
2001-12-04
期刊：
影响因子：
5
通讯作者：
Blaho, JA
中科院分区：
文献类型：
--
作者：
Blouin, A;Blaho, JA
我们先前证明，单纯疱疹病毒1型（HSV-1）结构蛋白VP 22在感染早期存在于细胞质中，并在感染晚期迁移至细胞核并在细胞核中积累（J. Virol. 73（8）（1999）6769）。本研究的目的是记录在不存在其他病毒多肽的情况下VP 22在细胞中的行为。我们表征了各种间接免疫荧光样品制备条件对VP 22在细胞中定位的影响，并确定了以下内容。(i)用甲醛固定并用丙酮透化，保持了细胞中微管的结构，就像我们观察到的经典微管组织中心一样。(ii)单独的丙酮或甲醇不能完全固定细胞。(iii)Triton X-100降低了我们系统中微管蛋白的免疫荧光信号。(iv)VP 22主要存在于甲醛固定的细胞核中。根据我们的研究结果，我们得出以下结论。(v)由于仅用丙酮或甲醇部分固定，微管形成弥散的不规则形状。(vi)仅在用丙酮或甲醇固定的细胞的细胞质中检测到VP 22，这是由于其从细胞核渗出。总之，这些发现表明（vii）VP 22的核定位不需要额外的病毒因子。(C)2001爱思唯尔科技有限公司。保留所有权利。
We previously demonstrated that the herpes simplex virus type 1 (HSV-1) structural protein VP22 exists in the cytoplasm early in infection and migrates to and accumulates in the nucleus late in infection (J. Virol. 73(8) (1999) 6769). The goal of this study is to document the behavior of VP22 in cells in the absence of other viral polypeptides. We characterized the effects of various indirect immunofluorescence sample preparation conditions on the localization of VP22 in cells and have determined the following. (i) Fixing with formaldehyde and permeabilizing with acetone maintains the structure of microtubules in cells, in as much as we observed classic microtubule organizing centers. (ii) Acetone or methanol alone did not completely fix the cells. (iii) Triton X-100 decreased tubulin immunofluorescence signals in our system. (iv) VP22 predominated in the nucleus of cells that were fixed with formaldehyde. Based on our results, we conclude the following. (v) Due to the partial fixation by acetone or methanol alone, microtubules form diffuse irregular shapes. (vi) VP22 is detected in the cytoplasm of cells fixed with acetone or methanol only due to its seepage from the nucleus. Taken together, these findings indicate that (vii) the nuclear localization of VP22 does not require additional viral factors. (C) 2001 Elsevier Science B.V. All rights reserved.