Identification, expression and subcellular localization of ESRG
Identification, expression and subcellular localization of ESRG
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ESRG的鉴定、表达和亚细胞定位
DOI：
10.1016/j.bbrc.2013.04.062
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发表时间：
2013-05-24
影响因子：
3.1
通讯作者：
Jiang, Xingjun
中科院分区：
文献类型：
--
作者：
Li, Guifei;Ren, Caiping;Jiang, Xingjun
胚胎干细胞相关基因（ESRG, embryonic stem cell related gene，简称HESRG）是本课组首次克隆并通过微阵列分析鉴定的人类新基因。有趣的是，它在未分化的人胚胎干细胞（hESCs）中特异性表达，但其表达模式及其在hESCs中的作用尚不清楚。本研究采用RNA连接酶介导的cDNA末端快速扩增技术（RLM-RACE）进一步鉴定全长3151nt ESRG cDNA。同时，还发现了一个长度为2837nt的选择性剪接ESRG转录物（ESRG- b）。令人惊讶的是，生物信息学分析表明，ESRG和ESRG- b的开放阅读框（orf）是相同的。它们都由669nt组成，编码一个222aa蛋白，预计分子大小为24 kDa。我们制备的ESRG特异性抗体经免疫细胞化学染色和Western blotting证实，ESRG蛋白位于hESCs的细胞核中。相比之下，ESRG通过基因转染策略在COS7细胞中被迫表达时位于细胞质中，这表明可能存在一些仅存在于hESCs中的特殊蛋白，可以帮助ESRG蛋白转运到hESCs的细胞核中。通过空间表达分析，我们进一步发现ESRG仅在卵巢组织和hESCs中表达，而不在其他组织或细胞系中表达。我们目前的数据为我们进一步研究ESRG在hESCs中的功能和调控机制提供了重要的基础。(C) 2013爱思唯尔公司版权所有。
ESRG (embryonic stem cell related gene, also known as HESRG), is a novel human gene first cloned and identified by our group with microarray analysis. Interestingly, it is expressed specifically in undifferentiated human embryonic stem cells (hESCs), while its expression pattern and its role in hESCs remain unclear. Here, full-length 3151nt ESRG cDNA was further identified by RNA ligase mediated rapid amplification of cDNA ends (RLM-RACE) technique. Meanwhile, an alternatively splicing ESRG transcript (ESRG-B) of 2837nt in length was also found. Surprisingly, bioinformatics analyses showed that the open reading frames (ORFs) of ESRG and ESRG-B were identical. Both of them consist of 669nt and encode a 222aa protein with a predicted molecular size of 24 kDa. The ESRG protein was located in the nuclei of hESCs as demonstrated by immunocytochemical staining and Western blotting using ESRG specific antibody generated by us. In contrast, ESRG located in the cytoplasm of COS7 cells when it was forced to be expressed in these cells by gene transfection strategy, suggesting there may be some special proteins present only in hESCs which can help ESRG protein transport into the nuclei of hESCs. By spatial expression analysis, we further discovered that ESRG only expressed in the ovary tissue and hESCs instead of other tissues or cell lines. Our current data provide us with an important basis for conducting further studies on the functions and regulatory mechanisms underlying the role of ESRG in hESCs. (C) 2013 Elsevier Inc. All rights reserved.