Characterization of the nuclear localization signal of high risk HPV16 E2 protein
Characterization of the nuclear localization signal of high risk HPV16 E2 protein
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DOI：
10.1016/j.virol.2006.10.018
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发表时间：
2007-03-30
期刊：
影响因子：
3.7
通讯作者：
Moroianu, Junona
中科院分区：
文献类型：
--
作者：
Klueevsek, Kristin;Wertz, Mary;Moroianu, Junona
高危人乳头状病毒 16 型 (HPV16) 的 E2 蛋白包含氨基末端 (N) 结构域、铰链 (H) 区域和羧基末端 (C) DNA 结合结构域。在 HeLa 细胞的转染测定中使用增强型绿色荧光蛋白 (EGFP) 与全长 E2 和 E2 结构域的融合，我们发现 C 结构域负责 E2 在体内的核定位，而 N 和 H 结构域不包含额外的核定位信号 (NLS)。 EGFP-E2 和 EGFP-cE2 的删除分析确定 C 结构域包含与 DNA 结合区域重叠的 a 螺旋 cNLS。突变分析表明，该 cNLS 中的精氨酸和赖氨酸残基对于 HPV16 E2 的核定位至关重要。有趣的是，这些碱性氨基酸残基在 BPV-1 和一些高风险 HPV 类型的 E2 蛋白中非常保守，但在低风险 HPV 类型中则不然，这表明这些 E2 蛋白之间的 NLS 和相应的核输入途径之间存在差异。 (c) 2006 Elsevier Inc. 保留所有权利。
The E2 protein of high risk human papillornavirus type 16 (HPV16) contains an amino-terminal (N) domain, a hinge (H) region and a carboxyl-tenrimal (C) DNA-binding domain. Using enhanced green fluorescent protein (EGFP) fusions with full length E2 and E2 domains in transfection assays in HeLa cells, we found that the C domain is responsible for the nuclear localization of E2 in vivo, whereas the N and H domains do not contain additional nuclear localization signals (NLSs). Deletion analysis of EGFP-E2 and EGFP-cE2 determined that the C domain contains an a helix cNLS that overlaps with the DNA-binding region. Mutational analysis revealed that the arginine and lysine residues in this cNLS are essential for nuclear localization of HPV16 E2. Interestingly, these basic amino acid residues are well conserved arnong the E2 proteins of BPV-1 and some high risk HPV types but not in the low risk HPV types, suggesting that there are differences between the NLSs and corresponding nuclear import pathways between these E2 proteins. (c) 2006 Elsevier Inc. All rights reserved.