Zhao Qingguo, Zhou Yu, Cao Zhiguo, Zhu Hengqi, Huang Peitang, Lu Baisong
Beijing Institute of Biotechnology, Beijing, China.
Mol Reprod Dev. 2005 Sep;72(1):31-9. doi: 10.1002/mrd.20313.
Gametogenetin (Ggn) is a germ cell-specific gene with multiple splicing variants giving rise to three predicted protein products, gametogenetin protein 1 (GGN1), gametogenetin protein 2 (GGN2), and gametogenetin protein 3 (GGN3). GGN1 and GGN3 were reported to interact with Fanconi anemia complementation group L (FANCL) per proliferation of germ cells (POG), a ubiquitin E3 ligase involved in germ-cell-deficient (gcd) mutation. While GGN2, another protein from Ggn by alternative splicing did not interact with FANCL/POG since it lacked the domain mediating the interaction. Little is known about the expression and function of GGN2. Here through Northern blotting experiment we showed that Ggn was mainly expressed in the testis but hardly detectable in the ovary or the somatic tissues. By preparing GGN2-specific antibody we showed that GGN2 was detectable and only detectable in the testis. By comparing the expression of Ggn mRNA and GGN2 protein in developing mouse testis, we showed that there was no evident delay of the translation of Ggn mRNA after their transcription. Both the subcellular localization study and the germ cell membrane protein fractionation implied that GGN2 associated with the intracellular membrane system. Co-fractionation on Superdex and yeast two-hybrids suggested that like GGN1, GGN2 was also a potential interaction partner of gametogenetin binding protein 1 (GGNBP1). Our data suggested that gametogenetin proteins were mainly involved in male germ cell development and GGN2 was also a possible interaction partner of GGNBP1. Like GGN1, GGN2 was also possibly involved in cell trafficking. The possible involvement of GGN2 in acrosome biogenesis was proposed.
配子生成素(Ggn)是一种生殖细胞特异性基因,具有多种剪接变体,可产生三种预测的蛋白质产物,即配子生成素蛋白1(GGN1)、配子生成素蛋白2(GGN2)和配子生成素蛋白3(GGN3)。据报道,GGN1和GGN3在生殖细胞增殖(POG)过程中与范可尼贫血互补组L(FANCL)相互作用,FANCL是一种参与生殖细胞缺陷(gcd)突变的泛素E3连接酶。而通过可变剪接产生的Ggn的另一种蛋白质GGN2不与FANCL/POG相互作用,因为它缺乏介导相互作用的结构域。关于GGN2的表达和功能知之甚少。在这里,通过Northern印迹实验,我们表明Ggn主要在睾丸中表达,但在卵巢或体细胞组织中几乎检测不到。通过制备GGN2特异性抗体,我们表明GGN2可检测到且仅在睾丸中可检测到。通过比较发育中小鼠睾丸中Ggn mRNA和GGN2蛋白的表达,我们表明Ggn mRNA转录后其翻译没有明显延迟。亚细胞定位研究和生殖细胞膜蛋白分级分离均表明GGN2与细胞内膜系统相关。在Superdex上的共分级分离和酵母双杂交表明,与GGN1一样,GGN2也是配子生成素结合蛋白1(GGNBP1)的潜在相互作用伙伴。我们的数据表明,配子生成素蛋白主要参与雄性生殖细胞发育,GGN2也是GGNBP1的可能相互作用伙伴。与GGN1一样,GGN2也可能参与细胞运输。有人提出GGN2可能参与顶体生物发生。