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非洲爪蟾卵母细胞中多聚腺苷酸化组蛋白mRNA的存在情况。

On the existence of polyadenylated histone mRNA in Xenopus laevis oocytes.

作者信息

Levenson R G, Marcu K B

出版信息

Cell. 1976 Oct;9(2):311-22. doi: 10.1016/0092-8674(76)90121-5.

Abstract

In a variety of systems, histone mRNA has been shown to lack poly(A) (Adesnik and Darnell, 1972; Grunstein et al., 1973). We have found, however, that in Xenopus laevis oocytes, poly (A)-containing mRNA codes for histones, in a wheat germ cell-free system, based on the following criteria: first, co-migration with authentic X. laevis oocyte histones on polyacrylamide gels; second, no detectable incorporation of tryptophan; third, differential incorporation of lysine and methionine into histone fraction H2A; fourth, resistance of histone fraction H2A to cleavage with cyanogen bromide; and fifth, correspondence of tryptic peptide maps of partially purified cell-free products with authentic X. laevis oocyte histone. RNA which directs the synthesis of histones in the cell-free system is retained on oligo(dT)-cellulose, even after denaturation in 80% DMSO at 70 degrees C, thereby demonstrating the covalent attachment of polyadenylic acid sequences to the mRNA. Poly (A)- RNA (7S-14S fraction) was also found to code for histones using the same criteria. We discuss the significance of the finding that X. laevis oocytes contain two classes of histone mRNA as well as the potential developmental implications of this observation.

摘要

在多种系统中,组蛋白mRNA已被证明缺乏聚腺苷酸(Adesnik和Darnell,1972年;Grunstein等人,1973年)。然而,我们发现,在非洲爪蟾卵母细胞中,基于以下标准,含聚腺苷酸的mRNA在无细胞小麦胚芽系统中编码组蛋白:第一,在聚丙烯酰胺凝胶上与真实的非洲爪蟾卵母细胞组蛋白共迁移;第二,未检测到色氨酸的掺入;第三,赖氨酸和蛋氨酸向组蛋白H2A组分的差异掺入;第四,组蛋白H2A组分对溴化氰裂解具有抗性;第五,部分纯化的无细胞产物的胰蛋白酶肽图与真实的非洲爪蟾卵母细胞组蛋白相符。即使在70℃下于80%二甲基亚砜中变性后,在无细胞系统中指导组蛋白合成的RNA仍保留在寡聚(dT)-纤维素上,从而证明聚腺苷酸序列与mRNA的共价连接。使用相同标准还发现,聚(A)-RNA(7S-14S组分)也编码组蛋白。我们讨论了非洲爪蟾卵母细胞含有两类组蛋白mRNA这一发现的意义以及这一观察结果可能的发育影响。

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