Schumacher J M, Lee K, Edelhoff S, Braun R E
Department of Genetics, University of Washington, Seattle 98195, USA.
J Cell Biol. 1995 May;129(4):1023-32. doi: 10.1083/jcb.129.4.1023.
Previous studies in transgenic mice have established the importance of the 3' untranslated region (UTR) of the spermatid-specific protamine-1 (Prm-1) mRNA in its translational control during male germ cell development. To clone genes that mediate the translational repression or activation of the Prm-1 mRNA, we screened cDNA expression libraries made with RNA from pachytene spermatocytes and round spermatids, with an RNA probe corresponding to the 3' UTR of Prm-1. We obtained six independent clones that encode Spnr, a spermatid perinuclear RNA-binding protein. Spnr is a 71-kD protein that contains two previously described RNA binding domains. The Spnr mRNA is expressed at high levels in the testis, ovary, and brain, and is present in multiple forms in those tissues. Immunolocalization of the Spnr protein within the testis shows that it is expressed exclusively in postmeiotic germ cells and that it is localized to the manchette, a spermatid-specific microtubular array. Although the Spnr protein is expressed too late to be directly involved in the translational repression of Prm-1 specifically, we suggest that the Spnr protein may be involved in other aspects of spermatid RNA metabolism, such as RNA transport or translational activation.
先前对转基因小鼠的研究已经证实,精子细胞特异性鱼精蛋白-1(Prm-1)mRNA的3'非翻译区(UTR)在雄性生殖细胞发育过程中的翻译控制中具有重要作用。为了克隆介导Prm-1 mRNA翻译抑制或激活的基因,我们用对应于Prm-1 3'UTR的RNA探针筛选了由粗线期精母细胞和圆形精子细胞的RNA构建的cDNA表达文库。我们获得了六个独立的克隆,它们编码Spnr,一种精子细胞核周RNA结合蛋白。Spnr是一种71-kD的蛋白质,包含两个先前描述的RNA结合结构域。Spnr mRNA在睾丸、卵巢和脑中高水平表达,并且在这些组织中以多种形式存在。睾丸内Spnr蛋白的免疫定位表明,它仅在减数分裂后生殖细胞中表达,并且定位于精子细胞特异性微管阵列——精子颈部。尽管Spnr蛋白表达得太晚,无法直接参与Prm-1的特异性翻译抑制,但我们认为Spnr蛋白可能参与精子细胞RNA代谢的其他方面,如RNA转运或翻译激活。