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  

SND1 protein co-localization with TIA-1 on stress granules under stress stimuli

作　　者：邵洁[1] 张兵兵[1] 赵猛[1] 周云丽[1] 任丽[1] SHAO Jie;ZHANG Bing-bing;ZHAO Meng;ZHOU Yun-li;REN Li(Department of Clinical Laboratory, Cancer Institute and Hospital, Tianjin Medical University, National Clinical Research)

机构地区：[1]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

出　　处：《天津医药》2017年第6期561-565,共5页Tianjin Medical Journal

基　　金：国家自然科学基金青年项目(31501091)

摘　　要：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Western Blotting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(SN domain)。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。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 domain.TIA-1still can be recruited on stress granules but a large amount of stress granules were reducedeven though the expression of SND1protein was decreased.And the transportation of SND1on stress granules was lagedbehind TIA-1under different-times of heat shocks.Conclusion SND1protein co-localizes with TIA-1on stress granules,and which co-regulates the cellular stress response under stress stimuli.

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 

分 类 号：R349.5[医药卫生—基础医学]