Mol Reprod Dev. 2013 Dec;80(12):988-99. doi: 10.1002/mrd.22262.
Mouse testis actin-like proteins 1 and 2 (mTact1 and mTact2), which are expressed in murine haploid germ cells, have been described previously. Here, we report the cloning and characterization of a third actin-like protein from rat, rat testis actin-like protein 3 (rTact3). The complete cDNA of the rTact3 gene was approximately 3.7 kb in length, and its corresponding amino acid sequence consisted of 1219 amino acids. The rTact3 gene lacks introns, similar to mTact1 and mTact2. The 356 C-terminal amino acids of rTact3 showed 43% homology with mTact1, whereas the 863 N-terminal amino acids did not show any significant homology. Northern blot analysis revealed that rTact3 mRNA was expressed only in adult rat testes and not during the prepubescent stage. In situ hybridization revealed that rTact3 was expressed exclusively during round and elongated spermatids maturation stages in rat testes. Immunohistochemical experiments using antibodies raised against a synthetic peptide showed that the expression of the rTact3 protein was also restricted in round and elongated spermatids, specifically in the head and acrosome of mature rat sperm. The 5′-flanking region of the mTact3 gene was found to contain a TATA-box motif as well as two putative CREB/c-Jun and five C/EBP motifs. mTact3 promoter activity was enhanced in a dose-dependent manner by the transfection of CREB, c-Jun, or C/EBP in NIH3T3 cells. These results suggest that Tact3 proteins might play an important role in rodent germ-cell development.
鼠睾丸肌动蛋白样蛋白 1 和 2(mTact1 和 mTact2),以前曾在小鼠单倍体生殖细胞中表达,已经被描述过。在这里,我们报道了来自大鼠的第三种肌动蛋白样蛋白,即大鼠睾丸肌动蛋白样蛋白 3(rTact3)的克隆和特性。rTact3 基因的完整 cDNA 约为 3.7kb,其相应的氨基酸序列由 1219 个氨基酸组成。rTact3 基因缺乏内含子,与 mTact1 和 mTact2 相似。rTact3 的 356 个 C 末端氨基酸与 mTact1 具有 43%的同源性,而 863 个 N 末端氨基酸没有显示任何显著的同源性。Northern blot 分析显示 rTact3 mRNA 仅在成年大鼠睾丸中表达,而在青春期前阶段不表达。原位杂交显示 rTact3 仅在大鼠睾丸的圆形和伸长精子细胞成熟阶段表达。使用针对合成肽的抗体进行免疫组织化学实验表明,rTact3 蛋白的表达也局限于圆形和伸长的精子细胞,特别是在成熟大鼠精子的头部和顶体中。发现 mTact3 基因的 5'侧翼区包含 TATA 盒基序以及两个推定的 CREB/c-Jun 和五个 C/EBP 基序。在 NIH3T3 细胞中,通过转染 CREB、c-Jun 或 C/EBP,mTact3 启动子活性呈剂量依赖性增强。这些结果表明,Tact3 蛋白可能在啮齿动物生殖细胞发育中发挥重要作用。