Sun Yingjie, Zhang Pin, Zheng Hang, Dong Luna, Tan Lei, Song Cuiping, Qiu Xusheng, Liao Ying, Meng Chunchun, Yu Shengqing, Ding Chan
Department of Avian Infectious Diseases, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Science, Shanghai 200241, China.
College of Animal Science and Technology, Shandong Agricultural University, Taian 271018, China.
J Vet Sci. 2018 Jan 31;19(1):3-12. doi: 10.4142/jvs.2018.19.1.3.
T-cell internal antigen-1 (TIA-1) has roles in regulating alternative pre-mRNA splicing, mRNA translation, and stress granule (SG) formation in human cells. As an evolutionarily conserved response to environmental stress, SGs have been reported in various species. However, SG formation in chicken cells and the role of chicken TIA-1 (cTIA-1) in SG assembly has not been elucidated. In the present study, we cloned cTIA-1 and showed that it facilitates the assembly of canonical SGs in both human and chicken cells. Overexpression of the chicken prion-related domain (cPRD) of cTIA-1 that bore an N-terminal green fluorescent protein (GFP) tag (pntGFP-cPRD) or Flag tag (pFlag-cPRD) induced the production of typical SGs. However, C-terminal GFP-tagged cPRD induced notably large cytoplasmic granules that were devoid of endogenous G3BP1 and remained stable when exposed to cycloheximide, indicating that these were not typical SGs, and that the pntGFP tag influences cPRD localization. Finally, endogenous cTIA-1 was recruited to SGs in chicken cells and tissues under environmental stress. Taken together, our study provide evidence that cTIA-1 has a role in canonical SG formation in chicken cells and tissues. Our results also indicate that cPRD is necessary for SG aggregation.
T细胞内抗原1(TIA-1)在调节人类细胞中的可变前体mRNA剪接、mRNA翻译和应激颗粒(SG)形成方面发挥作用。作为对环境应激的一种进化保守反应,应激颗粒已在各种物种中被报道。然而,鸡细胞中的应激颗粒形成以及鸡TIA-1(cTIA-1)在应激颗粒组装中的作用尚未阐明。在本研究中,我们克隆了cTIA-1,并表明它促进了人类和鸡细胞中典型应激颗粒的组装。带有N端绿色荧光蛋白(GFP)标签(pntGFP-cPRD)或Flag标签(pFlag-cPRD)的cTIA-1的鸡朊病毒相关结构域(cPRD)的过表达诱导了典型应激颗粒的产生。然而,C端GFP标记的cPRD诱导出明显较大的细胞质颗粒,这些颗粒缺乏内源性G3BP1,并且在暴露于环己酰亚胺时保持稳定,这表明这些不是典型的应激颗粒,并且pntGFP标签影响cPRD的定位。最后,在环境应激下,内源性cTIA-1被招募到鸡细胞和组织中的应激颗粒中。综上所述,我们的研究提供了证据表明cTIA-1在鸡细胞和组织中的典型应激颗粒形成中发挥作用。我们的结果还表明cPRD是应激颗粒聚集所必需的。