天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (6): 561-565.doi: 10.11958/20170286

• 细胞与分子生物学 •    下一篇

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

邵洁，张兵兵，赵猛，周云丽，任丽△ #br# #br#   

  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
  • 收稿日期:2017-03-07 修回日期:2017-04-26 出版日期:2017-06-15 发布日期:2017-07-05
  • 通讯作者: △通讯作者 E-mail: roland-li@163.com E-mail:jieshao@tmu.edu.cn
  • 作者简介:邵洁（1980），女，博士研究生，讲师，主要从事免疫学及临床肿瘤标志物研究
  • 基金资助:
    国家自然科学基金青年项目（31501091）

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

SHAO Jie, ZHANG Bing-bing, ZHAO Meng, ZHOU Yun-li, REN Li   

  1. Department of Clinical Laboratory, Cancer Institute and Hospital, Tianjin Medical University, National Clinical Research Center of Cancer, Tianjin Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, China
  • Received:2017-03-07 Revised:2017-04-26 Published:2017-06-15 Online:2017-07-05

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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。

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

Abstract: Objective To analyze the association of staphylococcal nuclease domain-containing protein 1(SND1) and T-cell intracellular antigen 1(TIA-1) on stress granules, and the regulation of SND1 on stress granules under stress stimuli. Methods The immunofluorescence assay and laser scanning confocal microscopy were used to observe the co-localization of SND1 protein and TIA-1 protein under stress stimuli, and the over-expression plasmids of pEGFP vector were transfected into HeLa cells and to verify which domain of SND1 co- localized with TIA- 1 under stress stimuli. RNA interferencemediated knockdown of the expression of SND1 protein in HeLa cells was measured by Western Blotting assay. Then whether the knockdown of SND1 affected the recruitment of TIA- 1 on stress granules was observed. Heat shocks under different times were used to identify whether there were dynamic changes in transportation of SND1 and TIA-1 on stress granules. Results SND1 co-localized with TIA- 1 on stress granules under stress stimuli, and the associated domain of SND1 were SN domain. TIA-1 still can be recruited on stress granules but a large amount of stress granules were reduced even though the expression of SND1 protein was decreased. And the transportation of SND1 on stress granules was laged behind TIA-1 under different-times of heat shocks. Conclusion SND1 protein co-localizes with TIA-1 on stress granules, and which co-regulates the cellular stress response under stress stimuli.

Key words: stress, RNA interference, Staphylococcal nuclease domain- containing protein 1, T- cell intracellular antigen 1, stress granule