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ISSN 1672-9145 Acta Biochim Biophys Sin 2005, 37(6): 396–405 CN 31-1940/Q

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Molecular Cloning of a Novel Mouse Testis-specific Spermatogenic Cell Apoptosis Inhibitor Gene mTSARG7 as a Candidate Oncogene

Xiao-Jun TAN, Xiao-Wei XING, Lu-Yun LI, Zhao-Di WU, Chang-Gao ZHONG, Dong-Song NIE, Jun-Jiang FU, Yang XIANG, Yun DENG, and Guang-Xiu LU*

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

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.

Key words mTSARG7; testis; spermatogenesis; oncogene

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Received: January 10, 2005 Accepted: April 8, 2005

This work was supported by the grants from the National Natural Science Foundation of China (No. 30371493, No. G1999055901) and the Major State Basic Research Development Program of China (No. G1999055901)

*Corresponding author: Tel, 86-731-4373187; Fax, 86-731-4497661; E-mail, [email protected]