Characterization, expression pattern and chromosomal localization of the spermatogenesis associated 6 gene (Spata6).
Characterization, expression pattern and chromosomal localization of the spermatogenesis associated 6 gene (Spata6).
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DOI：
10.1093/molehr/gag047
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发表时间：
2003-06
影响因子：
4
通讯作者：
C. Oh;H. Aho;R. Shamsadin;K. Nayernia;C. Muller;U. Sancken;C. Szpirer;W. Engel;I. Adham
中科院分区：
文献类型：
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作者：
C. Oh;H. Aho;R. Shamsadin;K. Nayernia;C. Muller;U. Sancken;C. Szpirer;W. Engel;I. Adham
我们报告的精子发生相关基因6（Spata 6）编码的预测蛋白的488个氨基酸的克隆和鉴定。它与驱动蛋白相关蛋白的马达结构域和秀丽隐杆线虫神经钙传感器蛋白（NCS-2）具有相似性。该基因编码约2.6、约1.8和约1.2 kb的三种mRNA。2.6 kb的mRNA在睾丸、卵巢、胸腺和胎盘中低水平表达，而1.8和1.2 kb的转录物仅在睾丸中表达。1.8和1.2 kb转录本在单倍体生殖细胞中特异性表达。原位杂交结果表明Spata 6基因在精子细胞中的表达高于精母细胞和精原细胞。RT-PCR和原位杂交结果表明，Spata 6基因在小鼠胚胎发育过程中有表达，并定位于神经管、体节和肢芽中。Spata 6基因由15个外显子组成，大小在40和596 bp之间。2.6和1.8 kb转录物具有不同的5'非翻译序列，但具有相同的翻译起始位点，因此可能编码预测分子量为49.7 kDa的相同蛋白质。1.2kb转录物来源于外显子7和8之间的近端启动子，并且含有翻译起始密码子AUG，其与2.6和1.8kb转录物的起始密码子AUG在读框内。因此，1.2kb转录物可能编码32 kDa的截短蛋白。Western印迹分析与抗血清提出了对合成肽从C-末端的推导Spata 6蛋白检测只有一个单一的蛋白质的53 kDa的所有组织研究。Spata 6基因定位于大鼠5号染色体q34-35区域和人类1号染色体p32-35区域。为了确定Spata 6的功能，我们通过同源重组使胚胎干细胞中的小鼠基因失活。尽管杂合突变细胞能够产生低毛色嵌合小鼠，但所有嵌合体都没有将靶向等位基因传递给它们的后代，这表明Spata 6（+/-）细胞的高贡献导致嵌合胚胎的致死性。
We report the cloning and characterization of the spermatogenesis associated 6 gene (Spata6) encoding a predicted protein of 488 amino acids. It exhibits similarity with the motor domain of kinesin related proteins and with the Caenorhabditis elegans neural calcium sensor protein (NCS-2). The gene encodes three mRNAs of approximately 2.6, approximately 1.8 and approximately 1.2 kb. The expression of the 2.6 kb mRNA is detected at low levels in testis, ovary, thymus and placenta, while the 1.8 and 1.2 kb transcripts are exclusively expressed in testis. The 1.8 and 1.2 kb transcripts are specifically expressed in haploid germ cells. Data from in situ hybridization experiments suggested that mRNA expression of Spata6 in spermatids is higher than in spermatocytes and spermatogonia. RT-PCR analysis and whole mount in situ hybridization demonstrate that the Spata6 transcript is expressed during embryonic development and is localized in neural tube, somites and limb buds of mouse embryo. The Spata6 gene consists of 15 exons ranging in size between 40 and 596 bp. The 2.6 and 1.8 kb transcripts have different 5' untranslated sequences but have the same translational initiation site and therefore may encode the same protein with a predicted molecular weight of 49.7 kDa. The 1.2 kb transcript is derived from a proximal promoter between exons 7 and 8, and contains a translation initiation codon AUG, which is in frame with initiator AUG codon of the 2.6 and 1.8 kb transcripts. Therefore, the 1.2 kb transcript may code for a truncated protein of 32 kDa. Western blot analysis with the antiserum raised against a synthetic peptide from the C-terminal of the deduced Spata6 protein detects only a single protein of 53 kDa in all tissues studied. The Spata6 gene was localized to chromosome 5, region q34-35 in the rat and to chromosome 1, region p32-35 in the human. In an effort to determine the function of Spata6, we inactivated the mouse gene in embryonic stem cells through homologous recombination. Although the heterozygous mutant cells were able to generate low coat colour chimeric mice, all chimeras did not transmit the targeted allele to their progeny suggesting that a high contribution of Spata6(+/-) cells lead to the lethality of the chimeric embryos.