[1]李　鹏,刘　冰,王志向,等.肾癌转移相关基因CXC趋化因子受体4核定位信号的结构分析与功能鉴定[J].新乡医学院学报,2017,34(2):086-89.[doi:10.7683/xxyxyxb.2017.02.002]
　LI Peng,LIU Bing,WANG Zhi-xiang,et al.Structural analysis and functional identification of nuclear localization signal sequence of CXC chemokine receptor type 4[J].Journal of Xinxiang Medical University,2017,34(2):086-89.[doi:10.7683/xxyxyxb.2017.02.002]
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肾癌转移相关基因CXC趋化因子受体4核定位信号的结构分析与功能鉴定
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
34
期数:
2017年2
页码:
086-89
栏目:
基础研究
出版日期:
2017-02-05

文章信息/Info

Title:
Structural analysis and functional identification of nuclear localization signal sequence of CXC chemokine receptor type 4
作者:
李　鹏1; 2; 刘　冰1; 王志向1; 鲍　一1; 陈俊明1; 吴登爽1; 吴震杰1; 王林辉1
（1.第二军医大学附属长征医院泌尿外科，上海　200003；2.解放军第一五三中心医院泌尿外科，河南　郑州　450014）
Author(s):
LI Peng1; 2; LIU Bing1; WANG Zhi-xiang1; BAO Yi1; CHEN Jun-ming1; WU Deng-shuang1; WU Zhen-jie1; WANG Lin-hui1
（1.Department of Urinary Surgery，Changzheng Hospital Affiliated to the Second Military Medical University，Shanghai 200003，China；2.Department of Urinary Surgery，the No.153 Hospital of Chinese People′s Liberation Army，Zhengzhou 450014，Henan Province，China）
关键词:
肾细胞癌; CXC趋化因子受体4; 亚细胞分布; 核分布
Keywords:
renal cell carcinoma; CXC chemokine receptor type 4; subcellular distribution; nuclear localization
分类号:
R737.11
DOI:
10.7683/xxyxyxb.2017.02.002
文献标志码:
A
摘要:
目的　分析肾癌转移相关基因CXC趋化因子受体4（CXCR4）核定位信号序列的组成结构，并鉴定其功能。方法　首先通过生物信息学方法检索Ensembl和PSORTⅡ Prediction在线数据库，分析CXCR4核定位信号序列的组成和结构。然后根据生物信息学分析结果，通过重叠延伸聚合酶链反应和基因克隆技术构建含有CXCR4核定位信号序列突变基因与绿色荧光蛋白基因的融合基因质粒，并将其转染入肾癌细胞表达，在荧光和激光共聚焦显微镜下观察CXCR4核定位信号序列突变体蛋白的亚细胞分布情况。结果　CXCR4蛋白的核定位信号序列为其第146~149位氨基酸，组成为精氨酸-脯氨酸-精氨酸-赖氨酸（RPRK）。荧光和激光共聚焦显微镜下，核定位信号序列突变的CXCR4蛋白分布于肾癌细胞膜和细胞质，不再分布于细胞核，而野生型CXCR4蛋白除细胞膜和细胞质外，仍可分布于肾癌细胞核。结论　CXCR4蛋白的核定位信号序列确为其第146~149位氨基酸，该突变序列中任一碱性氨基酸位点均可导致CXCR4核分布能力丧失。
Abstract:
Objective　To analyze the structural composition of CXC chemokine receptor type 4 （CXCR4) nuclear localization signal(NLS) sequence，and identify its function.Methods　The structural composition of CXCR4 NLS was analyzed through online database of Ensembl and PSORT Ⅱ Prediction by bioinformatics retrieval，which the plasmids containing the fusion gene of mutated CXCR4 NLS and green fluorescent protein was established by overlapping extension polymerase chain reaction(PCR) and gene cloning techniques.Then the plasmids were transfected into the renal carcinoma cells in vitro.The subcellular distribution of the mutated CXCR4 protein，which was expressed in the renal carcinoma cells，was observed under the fluorescence microscope and the laser confocal microscope.Results　The 146th-149th amino acids in CXCR4 protein was its NLS，and its structural composition was arginine-proline-arginine-lysine (RPRK).Under the fluorescence and confocal microscopy，the mutated CXCR4 protein located in the cell membrane and cytoplasm of the renal cell carcinoma；but not distributed in the cell nuclear.The wild CXCR4 protein still distributed in the cell nuclear besides distributed in the cell membrane and cytoplasm of renal cell carcinoma.Conclusion　The 146th-149th amino acids is the NLS sequence of CXCR4.The mutation of NLS can result in the cell nuclear distribution disability of CXCR4 protein.

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更新日期/Last Update: 2017-02-05