A 110 nt fragment in the 5\'UTR of STAT2 that determines IRES activity and influences cell proliferation
首发时间:2021-04-07
Abstract:As a unique member of the STAT (Signal Transducer and Activator of Transcription) proteins family, STAT2 cannot recognize DNA target sites as homodimers like the others, yet it plays a critical role in type I interferon (IFN-I) signaling and many other signaling pathways. It was reported that STAT2 could help cells to cope with stress conditions by promoting antiviral responses in normal cells or drug resistance in carcinoma cells. As IRES (Internal Ribosome Entry Sites) were known to help promoting survival of cells under stress, together with the discovery that the 5\'UTR of STAT2 could form stable stem-loop secondary structures with a length of 203 nt, suggesting that there might be IRES activity. To testify this speculation, bicistronic reporter assay was designed as the main method to find out whether the 5\'-UTR of STAT2 harbors an IRES element or not. During the process, the IRES activity was confirmed, followed by the discovery that IRES active central domain was located at nt 33-142 in the 5\'UTR of STAT2. Afterwards, CRISPR/Cas9 techniques were utilized to construct a knockout cell line with the deletion of sequence between nt 65-129 in 5\'UTR. And it was discovered that knockout cells grew slightly faster than those wild type ones, which meant that the knocked-out fragment had an impact on normal cell proliferation. The exact mechanism behind this result remains to be further investigated.
keywords: Biochemistry and Molecular Biology 5\'UTR STAT2 IRES
点击查看论文中文信息
STAT2 5\'非翻译区中决定了IRES活性并能影响细胞增殖的一段110nt序列
摘要:作为信号转导及转录激活(Signal Transducer and Activator of Transcription,STAT)蛋白家族中独特的一员,STAT2无法像其它成员那样形成同源二聚体,但其仍然在I型干扰素与很多其它信号通路中扮演了关键角色。据报道,STAT2可以帮助细胞来应对压力环境,比如促进正常细胞的抗病毒反应或是提升癌细胞的抗药性。由于已知内部核糖体进入位点(Internal Ribosome Entry Sites,IRES)能够协助提升细胞在压力下的生存能力,并结合STAT2的5\'非翻译区(长度203 nt)能够形成稳定茎环二级结构的相关发现,表明了该区域可能具有IRES活性。为了验证这一假设,双顺反子报告试验被设计用于测试STAT2的5\'非翻译区是否含有IRES元件。在此过程中,STAT2的IRES活性得到了验证,紧接着IRES活性中心域被发现位于其 5\'非翻译区中的nt 33-142区域。接下来,CRISPR/Cas9技术被用于构建删除了5\'非翻译区nt 65-129序列的敲除细胞株。结果发现,敲除细胞的生长速度稍快于野生型细胞,这意味着敲除掉的片段对正常细胞增殖是有影响的。该现象背后的具体机制有待进一步调查。
关键词: 生物化学与分子生物学 5\'UTR STAT2 IRES
基金:
引用
No.****
动态公开评议
共计0人参与
勘误表
STAT2 5\'非翻译区中决定了IRES活性并能影响细胞增殖的一段110nt序列
评论
全部评论0/1000