Bidirectional increase in permeability of nuclear envelope upon poliovirus infection and accompanying alterations of nuclear pores
Bidirectional increase in permeability of nuclear envelope upon poliovirus infection and accompanying alterations of nuclear pores
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DOI：
10.1128/jvi.78.18.10166-10177.2004
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发表时间：
2004-09-01
影响因子：
5.4
通讯作者：
Agol, VI
中科院分区：
文献类型：
--
作者：
Belov, GA;Lidsky, PV;Agol, VI
脊髓灰质炎病毒和其他一些小核糖核酸病毒触发某些核蛋白重新定位到细胞质中。在这里，通过使用一种蛋白质改变其荧光颜色随时间的推移，并包含一个核定位信号（NLS），我们表明，脊髓灰质炎病毒引发的搬迁主要是由于退出的预合成的核蛋白进入细胞质。核膜的渗漏也通过毛地黄皂苷透化的病毒感染（但非模拟感染）细胞的细胞核不能保留含NLS的绿色荧光蛋白（GFP）衍生物来记录。在感染过程中，细胞质到细胞核的运输也得到了促进，如GAPDH（甘油醛-3-磷酸脱氢酶）、细胞周期蛋白B1和GFP的缺乏NLS的衍生物的实验所证明的，这些在未感染的细胞中主要是细胞质的。电子显微镜显示，在未感染细胞中观察到的核孔通道中的棒状屏障结构在感染细胞中缺失，给人以完全开放的孔的印象。脊髓灰质炎病毒2A蛋白酶的瞬时表达也导致核蛋白的重新定位。来自脊髓灰质炎病毒感染或2A表达细胞的裂解物诱导3 X EGFP-NLS从透化未感染细胞的细胞核流出。这种活性被弹性蛋白酶抑制剂弹性蛋白酶和N-（甲氧基琥珀酰基）-L-丙氨酰-L-丙氨酰-L-脯氨酰-L-缬氨酸氯甲基酮（已知也是脊髓灰质炎病毒蛋白酶2A抑制剂的药物）、半胱天冬酶抑制剂zVAD（OMe）、feldine和一些其他蛋白酶抑制剂抑制。这些数据表明，2A引起核外排，可能与zVAD（OMe）. fmk敏感蛋白酶合作。然而，脊髓灰质炎病毒感染促进细胞核蛋白流出，也在caspase-3和caspase-9缺陷，表明流出可能发生没有这些酶的参与。在受感染的细胞的核质交通改变的生物相关性进行了讨论。
Poliovirus and some other picornaviruses trigger relocation of certain nuclear proteins into the cytoplasm. Here, by using a protein changing its fluorescence color with time and containing a nuclear localization signal (NLS), we demonstrate that the poliovirus-triggered relocation is largely due to the exit of presynthesized nuclear protein into the cytoplasm. The leakiness of the nuclear envelope was also documented by the inability of nuclei from digitonin-permeabilized, virus-infected (but not mock-infected) cells to retain an NLS-containing derivative of green fluorescent protein (GFP). The cytoplasm-to-nucleus traffic was also facilitated during infection, as evidenced by experiments with GAPDH (glyceraldehyde-3-phosphate dehydrogenase), cyclin B1, and an NLS-lacking derivative of GFP, which are predominantly cytoplasmic in uninfected cells. Electron microscopy demonstrated that a bar-like barrier structure in the channel of the nuclear pores, seen in uninfected cells, was missing in the infected cells, giving the impression of fully open pores. Transient expression of poliovirus 2A protease also resulted in relocation of the nuclear proteins. Lysates from poliovirus-infected or 2A-expressing cells induced efflux of 3 X EGFP-NLS from the nuclei of permeabilized uninfected cells. This activity was inhibited by the elastase inhibitors elastatinal and N-(methoxysuccinyl)-L-alanyl-L-alanyl-L-prolyl-L-valine chloromethylketone (drugs known also to be inhibitors of poliovirus protease 2A), a caspase inhibitor zVAD(OMe), fmk, and some other protease inhibitors. These data suggest that 2A elicited nuclear efflux, possibly in cooperation with a zVAD(OMe).fmk-sensitive protease. However, poliovirus infection facilitated nuclear protein efflux also in cells deficient in caspase-3 and caspase-9, suggesting that the efflux may occur without the involvement of these enzymes. The biological relevance of nucleocytoplasmic traffic alterations in infected cells is discussed.