Dass B, McMahon K W, Jenkins N A, Gilbert D J, Copeland N G, MacDonald C C
Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, Lubbock, Texas 79430, USA.
J Biol Chem. 2001 Mar 16;276(11):8044-50. doi: 10.1074/jbc.M009091200. Epub 2000 Dec 11.
Many mRNAs in male germ cells lack the canonical AAUAAA but are normally polyadenylated (Wallace, A. M., Dass, B., Ravnik, S. E., Tonk, V., Jenkins, N. A., Gilbert, D. J., Copeland, N. G., and MacDonald, C. C. (1999) Proc. Natl. Acad Sci. U. S. A. 96, 6763-6768). Previously, we demonstrated the presence of two distinct forms of the M(r) 64,000 protein of the cleavage stimulation factor (CstF-64) in mouse male germ cells and in brain, a somatic M(r) 64,000 form and a variant M(r) 70,000 form. The variant form was specific to meiotic and postmeiotic germ cells. We localized the gene for the somatic CstF-64 to the X chromosome, which would be inactivated during male meiosis. This suggested that the variant CstF-64 was an autosomal homolog activated during that time. We have named the variant form "tau CstF-64," and we describe here the cloning and characterization of the mouse tauCstF-64 cDNA, which maps to chromosome 19. The mouse tauCstF-64 protein fits the criteria of the variant CstF-64, including antibody reactivity, size, germ cell expression, and a common proteolytic digest pattern with tauCstF-64 from testis. Features of mtauCstF-64 that might allow it to promote the germ cell pattern of polyadenylation include a Pro --> Ser substitution in the RNA-binding domain and significant changes in the region that interacts with CstF-77.
雄性生殖细胞中的许多mRNA缺乏典型的AAUAAA序列,但通常会进行多聚腺苷酸化(华莱士,A.M.,达斯,B.,拉夫尼克,S.E.,汤克,V.,詹金斯,N.A.,吉尔伯特,D.J.,科普兰,N.G.,以及麦克唐纳,C.C.(1999年)《美国国家科学院院刊》96卷,6763 - 6768页)。此前,我们证明在小鼠雄性生殖细胞和大脑中存在两种不同形式的切割刺激因子(CstF - 64)的64000分子量蛋白,一种是体细胞型的64000分子量形式,另一种是70000分子量的变体形式。变体形式特异于减数分裂和减数分裂后的生殖细胞。我们将体细胞型CstF - 64的基因定位到X染色体上,该染色体在雄性减数分裂期间会失活。这表明变体CstF - 64是在那时被激活的常染色体同源物。我们将变体形式命名为“tau CstF - 64”,在此描述小鼠tauCstF - 64 cDNA的克隆和特征,该cDNA定位于19号染色体。小鼠tauCstF - 64蛋白符合变体CstF - 64的标准,包括抗体反应性、大小、生殖细胞表达以及与睾丸中tauCstF - 64的共同蛋白水解消化模式。mtauCstF - 64可能促进多聚腺苷酸化的生殖细胞模式的特征包括RNA结合结构域中的脯氨酸→丝氨酸取代以及与CstF - 77相互作用区域的显著变化。