Cloning of a full-length cDNA of human testis-specific spermatogenic cell apoptosis inhibitor TSARG2 as a candidate oncogene.
A novel human gene full-length cDNA sequence-TSARG2 was identified from a human testis cDNA library using the SRG2 gene (GenBank Accession No. ), which was significantly up-regulated in cryptorchidism, as an electronic probe. TSARG2 was 1223 bp in length. The putative protein encoded by this gene was 305 amino acids with a theoretical molecular weight of 34,751 and isoelectric point of 9.85. The sequence shared no significant homology with any known protein in databases except SRG2. Northern blot analysis revealed that 1.7 kb TSARG2 transcript was detected selectively in human testis. Furthermore, results of in situ hybridization assay confirmed that TSARG2 was expressed in seminiferous tubules, more precisely in spermatogonia and spermatocyte. No mutation was found by PCR-SSCP in 122 cases of azoospermia, severe oligzoospermia, and cryptorchidism. The green fluorescence produced by pEGFP-C1/TSARG2 was detected on the nucleus of COS7 cells after 24h post-transfection. The pcDNA3.1(-)/TSARG2 plasmid was constructed and introduced into MCF7 cells by liposome transfection. TSARG2 can accelerate MCF7 cells to traverse the S-phase and enter the G2-phase compared with the control without transfection of TSARG2, which suggested that this gene plays an important role in the development of cryptorchid testis and is a testis-specific apoptosis candidate oncogene.
Liu
Shang-Feng
SF
Institute of Biomedicine, Tsinghua University, Beijing, People's Republic of China.
Lu
Guang-Xiu
GX
Liu
Gang
G
Xing
Xiao-Wei
XW
Li
Lu-Yun
LY
Wang
Zhao
Z
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D019556:COS Cells; D002455:Cell Division; D045744:Cell Line, Tumor; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D018899:CpG Islands; D017931:DNA Primers / Q000494:pharmacology; D018076:DNA, Complementary / Q000378:metabolism; D005091:Exons; D005260:Female; D015723:Gene Library; D049452:Green Fluorescent Proteins; D006801:Humans; D017403:In Situ Hybridization; D007438:Introns; D008164:Luminescent Proteins / Q000378:metabolism; D008297:Male; D008957:Models, Genetic; D008969:Molecular Sequence Data; D018807:Polymorphism, Single-Stranded Conformational; D011506:Proteins / Q000737:chemistry* / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012984:Software; D013737:Testis / Q000378:metabolism; D015854:Up-Regulation
D017931:DNA Primers; D018076:DNA, Complementary; D008164:Luminescent Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; C457219:SPATA4 protein, human; D049452:Green Fluorescent Proteins