Takamatsu N, Kanda H, Tsuchiya I, Yamada S, Ito M, Kabeno S, Shiba T, Yamashita S
Department of Biosciences, School of Science, Kitasato University, Kanagawa, Japan.
Mol Cell Biol. 1995 Jul;15(7):3759-66. doi: 10.1128/MCB.15.7.3759.
SRY-related cDNA encoding a protein with a high-mobility-group (HMG) box and a leucine zipper motif, which was designated SOX-LZ, was isolated from a rainbow trout testis cDNA library. Comparison of this cDNA with the mouse homologous cDNA isolated from a testis cDNA library exhibits an overall amino acid sequence identity of 77%, which is in striking contrast to the abrupt loss of amino acid sequence homology outside the HMG box found among mammalian SRY genes. In both rainbow trout and mice, Northern (RNA) blot analyses have revealed the presence of a testis-specific 3-kb-long SOX-LZ mRNA, and this transcript appeared coincidentally with the protamine mRNA, suggesting its expression in the germ line. A recombinant HMG box region protein encoded by SOX-LZ could bind strongly with an oligonucleotide containing an AACAAT sequence, which is also recognized by mouse Sry and Sox-5. Upon cotransfection into CHO cells, SOX-LZ transactivated transcription through its binding motif when the region including the leucine zipper motif was deleted [SOX-LZ (D105-356)]; however, the intact SOX-LZ failed to transactivate. The intact SOX-LZ could form homodimers through the leucine zipper, which resulted in inhibition of DNA binding by the HMG box, while SOX-LZ (D105-356), which was incapable of dimerization, showed specific binding with the AACAAT sequence. Thus, the repressed transactivation of the intact SOX-LZ in CHO cells was primarily attributable to the low level of DNA binding of SOX-LZ homodimers.
从虹鳟鱼睾丸cDNA文库中分离出一种与SRY相关的cDNA,其编码一种具有高迁移率族(HMG)盒和亮氨酸拉链基序的蛋白质,命名为SOX-LZ。将该cDNA与从小鼠睾丸cDNA文库中分离出的同源cDNA进行比较,发现其氨基酸序列总体同一性为77%,这与哺乳动物SRY基因中HMG盒外氨基酸序列同源性的突然丧失形成了鲜明对比。在虹鳟鱼和小鼠中,Northern(RNA)印迹分析均显示存在一种睾丸特异性的3kb长的SOX-LZ mRNA,并且该转录本与鱼精蛋白mRNA同时出现,表明其在生殖细胞系中表达。由SOX-LZ编码的重组HMG盒区域蛋白可与含有AACAAT序列的寡核苷酸强烈结合,该序列也能被小鼠Sry和Sox-5识别。当共转染到CHO细胞中时,当缺失包括亮氨酸拉链基序的区域[SOX-LZ(D105-356)]时,SOX-LZ通过其结合基序反式激活转录;然而,完整的SOX-LZ未能反式激活。完整的SOX-LZ可通过亮氨酸拉链形成同源二聚体,这导致HMG盒对DNA的结合受到抑制,而无法形成二聚体的SOX-LZ(D105-356)则与AACAAT序列表现出特异性结合。因此,完整的SOX-LZ在CHO细胞中反式激活受到抑制主要归因于SOX-LZ同源二聚体对DNA的低水平结合。