Nuclear/nucleolar localization properties of C-terminal nucleocapsid protein of SARS coronavirus.
Nuclear/nucleolar localization properties of C-terminal nucleocapsid protein of SARS coronavirus.
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DOI：
10.1016/j.virusres.2005.05.007
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发表时间：
2005-12
期刊：
影响因子：
5
通讯作者：
Zhu Y
中科院分区：
文献类型：
--
作者：
Timani KA;Liao Q;Ye L;Zeng Y;Liu J;Zheng Y;Ye L;Yang X;Lingbao K;Gao J;Zhu Y
一种新型冠状病毒（CoV）最近被确定为严重急性呼吸综合征（SARS）的病原体。冠状病毒科的核衣壳（N）蛋白没有可辨别的同源性，但它们具有共同的核仁-胞质分布模式。在N.为了确定这些假定的NLS在SARS-CoV N细胞内定位中的作用，我们使用兔抗N抗血清进行了共聚焦显微镜分析。在本报告中，我们表明，野生型N主要分布在细胞质中。N的N-末端含有NLS 1（aa 38 - 44），定位于细胞核。含有NLS 2（aa 257 - 265）和NLS 3（aa 369 - 390）的N的C-末端定位于细胞质和核仁。从各种缺失突变的分析结果表明，含有氨基酸226 - 289的区域能够介导核仁定位。缺失NLS 3两侧的两个疏水区域恢复了其活性并定位于细胞核。此外，亮氨酸富集区（220-LALLLLDRLNRL）的缺失导致N积累到细胞质和细胞核，当将该肽融合到EGFP时，定位于细胞质，这表明N可能充当穿梭蛋白。与其他冠状病毒家族成员相比，N的核/核仁定位特性的差异表明N具有独特的功能，这可能在SARS的发病机制中起重要作用。
A novel coronavirus (CoV) has recently been identified as the aetiological agent of severe acute respiratory syndrome (SARS). Nucleocapsid (N) proteins of the Coronaviridae family have no discernable homology, but they share a common nucleolar-cytoplasmic distribution pattern. There are three putative nuclear localization signal (NLS) motifs present in the N. To determine the role of these putative NLSs in the intracellular localization of the SARS–CoV N, we performed a confocal microscopy analysis using rabbit anti-N antisera. In this report, we show that the wild type N was distributed mainly in the cytoplasm. The N-terminal of the N, which contains the NLS1 (aa38–44), was localized to the nucleus. The C-terminus of the N, which contains both NLS2 (aa257–265) and NLS3 (aa369–390) was localized to the cytoplasm and the nucleolus. Results derived from analysis of various deletion mutations show that the region containing amino acids 226–289 is able to mediate nucleolar localization. The deletion of two hydrophobic regions that flanked the NLS3 recovered its activity and localized to the nucleus. Furthermore, deletion of leucine rich region (220-LALLLLDRLNRL) resulted in the accumulation of N to the cytoplasm and nucleolus, and when fusing this peptide to EGFP localization was cytoplasmic, suggesting that the N may act as a shuttle protein. Differences in nuclear/nucleolar localization properties of N from other members of coronavirus family suggest a unique function for N, which may play an important role in the pathogenesis of SARS.
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DOI：
10.1016/j.bbrc.2004.05.107
发表时间：
2004-07-09
影响因子：
3.1
作者：
Mizutani T;Fukushi S;Saijo M;Kurane I;Morikawa S
通讯作者：
Morikawa S
影响因子：
5
作者：
Hiscox JA
通讯作者：
Hiscox JA
影响因子：
5.4
作者：
Chen, CJ;Makino, S
通讯作者：
Makino, S
影响因子：
3.7
作者：
Mizutani T;Fukushi S;Saijo M;Kurane I;Morikawa S
通讯作者：
Morikawa S
影响因子：
3.8
作者：
DAVIES, HA;DOURMASHKIN, RR;MACNAUGHTON, MR
通讯作者：
MACNAUGHTON, MR