Identification of a novel isoform of DHRS4 protein with a nuclear localization signal.
Identification of a novel isoform of DHRS4 protein with a nuclear localization signal.
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DOI：
10.1016/j.gene.2011.12.033
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发表时间：
2012-02
期刊：
影响因子：
3.5
通讯作者：
Z. Su;Rui Li;Xuhong Song;Gefei Liu;Yifan Li;X. Chang;Cong-zhu Li;Dongyang Huang
中科院分区：
文献类型：
--
作者：
Z. Su;Rui Li;Xuhong Song;Gefei Liu;Yifan Li;X. Chang;Cong-zhu Li;Dongyang Huang
DHRS4基因编码NADP(H)依赖性视黄醇脱氢酶/还原酶(NRDR)，在调节视黄酸的合成中发挥重要作用。在本研究中，我们通过 RT-PCR、3' RACE 和 5' RACE 鉴定了人类 DHRS4 基因的一种新的剪接 RNA 变体，命名为 NRDRA2。 NRDRA2 mRNA 缺乏外显子 4 和 6，并且与 DHRS4 mRNA 相比，阅读框发生变化，导致 NRDR 过氧化物酶体靶向信号丢失，并在预测的 NRDRA2 蛋白中获得核定位信号。通过免疫沉淀和质谱分析鉴定了人宫颈癌细胞系 HeLa 中的内源性 NRDRA2 蛋白。绿色荧光蛋白报告基因检测显示NRDRA2蛋白主要定位于细胞核，证实其C末端的序列是合法的核定位信号序列。这项研究鉴定了编码亚细胞核定位 NRDRA2 蛋白的替代转录变体 NRDRA2。
The DHRS4 gene encodes an NADP(H)-dependent retinol dehydrogenase/reductase (NRDR) and plays an important role in regulating the synthesis of retinoic acid. In the present study, we identified a novel splice RNA variant, designated NRDRA2, of the human DHRS4 gene by RT-PCR, 3′ RACE, and 5′ RACE. NRDRA2 mRNA lacked exons 4 and 6, and had a shift in the reading frame when compared to DHRS4 mRNA, resulting in loss of the peroxisomal targeting signal of NRDR and gain of a nuclear localization signal in the predicted NRDRA2 protein. Endogenous NRDRA2 protein was identified in the human cervical carcinoma cell line HeLa by immunoprecipitation and mass spectrometric assay. A green fluorescent protein reporter assay showed that NRDRA2 protein mainly localized to the nuclei, confirming the sequence at its C-terminus as a legitimate nuclear localization signal sequence. This study identifies the alternative transcript variant NRDRA2 encoding a subcellular nuclear localized NRDRA2 protein.