Cellular uptake and nuclear delivery of recombinant adenovirus Penton base
Cellular uptake and nuclear delivery of recombinant adenovirus Penton base
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DOI：
10.1006/viro.1999.9864
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发表时间：
1999-09-15
期刊：
影响因子：
3.7
通讯作者：
Boulanger, P
中科院分区：
文献类型：
--
作者：
Hong, SS;Gay, B;Boulanger, P
发现表达为重组蛋白的Ad 2衣壳组分五邻体碱基能够影响HeLa细胞中腺病毒粒子的整个进入途径，即，细胞附着、胞吞作用、囊泡逃逸、胞质内运动和通过核孔复合体的移位。五聚化缺陷突变体的数据表明，这些连续的步骤都不依赖于五邻体基础五聚体状态，表明负责这些功能的肽结构域由单体携带。野生型（WT）和整合素结合位点双突变体（K288 E340）进行的观察表明，五邻体碱基可以进入细胞通过替代，RGD和LDV独立的途径。在昆虫细胞中发现的三个核寻址缺陷突变体中，只有一个，W165 H，在HeLa细胞中的核运输也发生了变化。另外两个，W119 H和RRR 547 EQQ，在HeLa细胞中显示核定位的WT模式，表明包括色氨酸-119和位置547处的基本信号的区域在人类细胞环境中不作为核定位信号。细胞结构和细胞骨架的完整性似乎是需要的矢量运动和核输入WT五邻体基地，所建议的实验，使用透化HeLa细胞，分离核膜，和细胞因子靶向药物。(C)北京：科学出版社.
An Ad2 capsid component, the penton base, expressed as recombinant protein, was found to be capable of affecting the entire entry pathway of adenovirion in HeLa cells, i.e., cell attachment, endocytosis, vesicular escape, intracytoplasmic movement, and translocation through the nuclear pore complex. Data with pentamerization-defective mutants suggested that none of these successive steps depended upon penton base pentamer status, indicating that the peptide domains responsible for these functions were carried by the monomer. Observations performed with wild-type (WT) and an integrin-binding-site double-mutant (K288E340) suggested that the penton base could enter the cell via an alternative, RGD- and LDV-independent, pathway. Of three mutants that were found to be defective in nuclear addressing in insect cells, only one, W165H, was also altered in nuclear transport in HeLa cells. The other two, W119H and RRR547EQQ, showed a WT pattern of nuclear localization in HeLa cells, suggesting that the region including tryptophan-119 and the basic signal at position 547 did not act as a nuclear localization signal in the human cell context. The integrity of cellular structures and the cytoskeleton seemed to be required for the vectorial movement and nuclear import of WT penton base, as suggested by experiments using permeabilized HeLa cells, isolated nuclear membranes, and cytoskeleton-targeted drugs. (C) 1999 Academic Press.