葡萄球菌核酸酶样结构蛋白1 在应激刺激下与T 细胞胞内抗原1 共同参与应激颗粒聚集

目的探讨葡萄球菌核酸酶样结构蛋白1（SND1）在应激刺激下如何与T细胞胞内抗原1（TIA-1）共同参与应激颗粒（SG）的聚集以及如何调节应激反应。方法利用免疫荧光实验和激光共聚焦显微镜观察HeLa细胞中的SND1蛋白与TIA-1蛋白在应激刺激下是否形成共定位颗粒，并利用绿色荧光蛋白载体过表达质粒转染HeLa细胞进行外源蛋白表达，从而确定在应激刺激下SND1蛋白与TIA-1结合的结构域。利用RNA干扰技术敲除HeLa细胞中SND1蛋白表达并利用WesternBlotting检测蛋白表达水平，从而观察SND1低表达是否对TIA-1聚集形成SG产生影响。利用不同热休克刺激时间观察SND1与TIA-1...

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Bibliographic Details
Published in天津医药 Vol. 45; no. 6; pp. 561 - 565
Main Author 邵洁;张兵兵;赵猛;周云丽;任丽
Format Journal Article
LanguageChinese
Published 天津医科大学肿瘤医院检验科,国家肿瘤临床医学研究中心,天津市恶性肿瘤临床医学研究中心,天津市"肿瘤防治"重点实验室 300060 2017
Subjects
干扰;葡萄球菌核酸酶样结构蛋白1;T
应激;RNA
细胞胞内抗原1;应激颗粒
RNA干扰
stress
T-cell intracellular antigen 1
T细胞胞内抗原1
应激颗粒
RNA interference
stress granule
应激
Staphylococcal nuclease domain-containing protein 1
葡萄球菌核酸酶样结构蛋白1
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ISSN0253-9896
DOI10.11958/20170286

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Summary:目的探讨葡萄球菌核酸酶样结构蛋白1（SND1）在应激刺激下如何与T细胞胞内抗原1（TIA-1）共同参与应激颗粒（SG）的聚集以及如何调节应激反应。方法利用免疫荧光实验和激光共聚焦显微镜观察HeLa细胞中的SND1蛋白与TIA-1蛋白在应激刺激下是否形成共定位颗粒，并利用绿色荧光蛋白载体过表达质粒转染HeLa细胞进行外源蛋白表达，从而确定在应激刺激下SND1蛋白与TIA-1结合的结构域。利用RNA干扰技术敲除HeLa细胞中SND1蛋白表达并利用WesternBlotting检测蛋白表达水平，从而观察SND1低表达是否对TIA-1聚集形成SG产生影响。利用不同热休克刺激时间观察SND1与TIA-1的聚集过程是否存在动态变化。结果SND1蛋白在应激刺激下与TIA-1蛋白结合共同参与SG聚集，其主要作用结构域为葡萄球菌核酸酶结构域（SNdomain）。SND1低表达不会抑制TIA-1聚集到SG，但会减少SG的数量。在不同热休克刺激时间下，SND1聚集到SG的运输过程滞后于TIA-1。结论SND1蛋白在应激刺激下与TIA-1蛋白共同参与SG的聚集，从而调节细胞应激反应。关键词：应激；RNA干扰；葡萄球菌核酸酶样结构蛋白1；T细胞胞内抗原1；应激颗粒
Bibliography:Objective To analyze the association of staphylococcal nuclease domain-containing protein1(SND1)andT-cell intracellular antigen1(TIA-1)on stress granules,and the regulation of SND1on stress granules under stress stimuli.Methods The immunofluorescence assay and laser scanning confocal microscopy were used to observe the co-localizationof SND1protein and TIA-1protein under stress stimuli,and the over-expression plasmids of pEGFP vector were transfectedinto HeLa cells and to verify which domain of SND1co-localized with TIA-1under stress stimuli.RNA interferencemediatedknockdown of the expression of SND1protein in HeLa cells was measured by Western Blotting assay.Thenwhether the knockdown of SND1affected the recruitment of TIA-1on stress granules was observed.Heat shocks underdifferent times were used to identify whether there were dynamic changes in transportation of SND1and TIA-1on stressgranules.Results SND1co-localized with TIA-1on stress granules under stress stimuli,and the associated domain ofSND1were SN d
ISSN:0253-9896
DOI:10.11958/20170286