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一种新型小鼠睾丸特异性生精细胞凋亡抑制基因mTSARG7作为候选癌基因的分子克隆

Molecular cloning of a novel mouse testis-specific spermatogenic cell apoptosis inhibitor gene mTSARG7 as a candidate oncogene.

作者信息

Tan Xiao-Jun, Xing Xiao-Wei, Li Lu-Yun, Wu Zhao-Di, Zhong Chang-Gao, Nie Dong-Song, Fu Jun-Jiang, Xiang Yang, Deng Yun, Lu Guang-Xiu

机构信息

Institute of Human Reproductive and Stem cell Engineering, Central South University, Changsha 410078, China.

出版信息

Acta Biochim Biophys Sin (Shanghai). 2005 Jun;37(6):396-405. doi: 10.1111/j.1745-7270.2005.00057.x.

Abstract

A novel mouse gene, mTSARG7 (GenBank accession No. AY489184), with a full cDNA length of 2279 bp and containing 12 exons and 11 introns, was cloned from a mouse expressed sequence tag (GenBank accession No. BE644543) that was significantly up-regulated in cryptorchidism. The gene was located in mouse chromosome 8A1.3 and encoded a protein containing 403 amino acid residues that was a new member of the acyltransferase family because the sequence contained the highly conserved phosphate acyltransferase (PlsC) domain existing in all acyltransferase-like proteins. The mTSARG7 protein and AU041707 protein shared 83.9% identity in 402 amino acid residues. Expression of the mTSARG7 gene was restricted to the mouse testis. The results of the in situ hybridization analysis revealed that the mTSARG7 mRNA was expressed in mouse spermatogonia and spermatocytes. Subcellular localization studies showed that the EGFP-tagged mTSARG7 protein was localized in the cytoplasm of GC-1 spg cells. The mTSARG7 mRNA expression was initiated in the mouse testis in the second week after birth, and the expression level increased steadily with spermatogenesis and sexual maturation of the mouse. The results of the heat stress experiment showed that the mTSARG7 mRNA expression gradually decreased as the heating duration increased. The pcDNA3.1 Hygro(-)/mTSARG7 plasmid was constructed and introduced into GC-1 spg cells by liposome transfection. The mTSARG7 can accelerate GC-1 spg cells, causing them to traverse the S-phase and enter the G2-phase, compared with the control group where this did not occur as there was no transfection of mTSARG7. In conclusion, our results suggest that this gene may play an important role in spermatogenesis and the development of cryptorchid testes, and is a testis-specific apoptosis candidate oncogene.

摘要

一个新的小鼠基因mTSARG7(GenBank登录号:AY489184),其cDNA全长2279 bp,包含12个外显子和11个内含子,是从一个在隐睾症中显著上调的小鼠表达序列标签(GenBank登录号:BE644543)克隆而来。该基因位于小鼠8A1.3染色体上,编码一个含有403个氨基酸残基的蛋白质,它是酰基转移酶家族的一个新成员,因为该序列包含在所有酰基转移酶样蛋白中都存在的高度保守的磷酸酰基转移酶(PlsC)结构域。mTSARG7蛋白与AU041707蛋白在402个氨基酸残基上具有83.9%的同一性。mTSARG7基因的表达局限于小鼠睾丸。原位杂交分析结果显示,mTSARG7 mRNA在小鼠精原细胞和精母细胞中表达。亚细胞定位研究表明,EGFP标记的mTSARG7蛋白定位于GC-1 spg细胞的细胞质中。mTSARG7 mRNA表达在出生后第二周开始于小鼠睾丸,并且随着小鼠的精子发生和性成熟,表达水平稳步增加。热应激实验结果表明,随着加热时间的增加,mTSARG7 mRNA表达逐渐降低。构建了pcDNA3.1 Hygro(-)/mTSARG7质粒,并通过脂质体转染将其导入GC-1 spg细胞。与未转染mTSARG7因而未出现这种情况的对照组相比,mTSARG7可加速GC-1 spg细胞,使其穿过S期并进入G2期。总之,我们的结果表明,该基因可能在精子发生和隐睾睾丸的发育中起重要作用,并且是一个睾丸特异性凋亡候选癌基因。

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