Molecular cloning and functional expression of nucleolar phospholipid hydroperoxide glutathione peroxidase in mammalian cells
Molecular cloning and functional expression of nucleolar phospholipid hydroperoxide glutathione peroxidase in mammalian cells
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DOI：
10.1016/j.bbrc.2003.09.183
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发表时间：
2003-11-07
影响因子：
3.1
通讯作者：
Nakagawa, Y
中科院分区：
文献类型：
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作者：
Nakamura, T;Imai, H;Nakagawa, Y
我们从大鼠和小鼠睾丸中克隆了磷脂氢谷胱甘肽过氧化物酶（PHGPx）的全长cDNA，包括外显子Ib。大鼠核PHGPx的N端的核信号序列具有不同于先前报道的大鼠精子核GPx（SnGPx）的序列。这种PHGPx-YFP（黄色荧光蛋白）融合蛋白的表达，包括一个新的核信号序列是专门定位在核仁中，虽然YFP融合只有一个新的核信号序列分布在整个细胞核中，表明优先转运核仁PHGPx到核仁所需的核信号序列和内部序列的PHGPx。核仁PHGPx在几种组织中低水平表达，但在睾丸中广泛表达。免疫组织化学分析表明，核仁PHGPx的表达在精原细胞，精母细胞和精子细胞的核仁。在RBL 2 H3细胞中过表达34 kDa核仁PHGPx显著抑制由放线菌素D和多柔比星诱导的细胞死亡，所述细胞死亡诱导核仁损伤。这些结果表明，核仁PHGPx在哺乳动物细胞防止核仁损伤中起重要作用。(C)2003年爱思唯尔公司All rights reserved.
We cloned a full-length cDNA for phospholipid hydroperoxide glutathione peroxidase (PHGPx) including exon Ib from rat and mouse testis. The nuclear signal sequence of the N terminal of rat nuclear PHGPx possessed a different sequence from that previously reported for rat sperm nuclei GPx (SnGPx). Expression of this PHGPx-YFP (yellow fluorescent protein) fusion protein including a novel nuclear signal sequence was exclusively localized in nucleolus; although YFPs fused with only a novel nuclear signal sequence were distributed in the whole nucleus, indicating that preferential translocation of nucleolar PHGPx into nucleoli was required for the nuclear signal sequence and internal sequence of PHGPx. Low level expression of nucleolar PHGPx was detected in several tissues, but the expression of nucleolar PHGPx was extensively high in testis. Immunohistochemical analysis with anti-nucleolar PHGPx indicated that expression of nucleolar PHGPx was observed in the nucleoli in the spermatogonia, spermatocyte, and spermatid. Overexpression of 34 kDa nucleolar PHGPx in RBL2H3 cells significantly suppressed cell death induced by actinomycin D and doxorubicin that induced damage in the nucleolus. These results indicated that nucleolar PHGPx plays an important role in prevention of nucleolus from damage in mammalian cells. (C) 2003 Elsevier Inc. All rights reserved.