Molecular cloning and identification of mouse epididymis-specific gene mHong1, the homologue of rat HongrES1
Molecular cloning and identification of mouse epididymis-specific gene mHong1, the homologue of rat HongrES1
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DOI：
10.1038/aja.2011.176
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发表时间：
2012-07-01
影响因子：
2.9
通讯作者：
Zhang, Yong-Lian
中科院分区：
文献类型：
--
作者：
Hu, Shuang-Gang;Du, Han;Zhang, Yong-Lian
已有研究表明，大鼠附睾特异性基因HongrES 1在精子获能和生育力中起重要作用。本研究通过序列比对和RT-PCR方法克隆了小鼠同源基因mHong 1。mHong 1基因位于染色体12 p14，跨越5个外显子。该cDNA序列由1257个核苷酸组成，编码一个419个氨基酸的蛋白质，预测N端信号肽为20个氨基酸。mHong 1 mRNA在表达模式上与HongrES 1相似：（i）在附睾组织中特异性表达，特别是在尾部区域;（ii）雄激素依赖性但睾丸流体因子独立性。其蛋白产物与HongrES 1具有71%的相似性，并且与HongrES 1一样含有经典的丝氨酸蛋白酶抑制剂结构域。制备了特异性和敏感性均较高的抗mHong 1多克隆抗体。与HongrES 1一样，mHong 1蛋白在附睾上皮中显示棋盘式表达模式，并分泌到附睾腔中。mHong 1蛋白显示出比HongrES 1更高的糖基化。虽然它们都沉积在精子头部表面，但mHong 1定位于赤道段，这与HongrES 1不同。mHong 1蛋白可以通过高离子强度从精子膜上去除，因此可以归类为外源性膜蛋白。综上所述，我们认为mHong 1是HongrES 1的同源物，本研究为建立动物模型阐明HongrES 1和mHong 1的确切功能奠定了基础。Asian Journal of Andrology（2012）14，626-634; doi：10.1038/aja.2011.176; 2012年3月19日在线发表
Previous studies have shown that rat epididymis-specific gene HongrES1 plays important roles in sperm capacitation and fertility. In this study, we cloned the mouse homologue gene by sequence alignment and RT-PCR methods and designated it as mHong1. The mHong1 gene is located on chromosome 12p14, spanning five exons. The cDNA sequence consists of 1257 nucleotides and encodes a 419 amino-acid protein with a predicted N-terminal signal peptide of 20 amino acids. The mHong1 mRNA shows similarity with HongrES1 in the expression patterns: (i) specific expression in epididymal tissue, especially in the cauda region; and (ii) androgen-dependence but testicular fluid factor independence. Its protein product shows 71% similarity with HongrES1 and contains a classical serpin domain as does HongrES1. A polyclonal antibody against mHong1 with high specificity and sensitivity was raised. Like HongrES1, the mHong1 protein shows a checker-board expression pattern in the epididymal epithelium and is secreted into the epididymal lumen. The mHong1 protein shows higher glycosylation than HongrES1. Although both of them are deposited onto the sperm head surface, mHong1 is localized to the equatorial segment, which is different from that of HongrES1. The mHong1 protein can be removed from the sperm membrane by high ionic strength and therefore can be classed as an extrinsic membrane protein. Collectively, we conclude that mHong1 is the homologue of HongrES1 and the present work paves the way for establishing animal models to elucidate the precise functions of HongrES1 and mHong1. Asian Journal of Andrology (2012) 14, 626-634; doi:10.1038/aja.2011.176; published online 19 March 2012