Nadin-Davis S, Mezl V A
Can J Biochem Cell Biol. 1983 Jun;61(6):353-62. doi: 10.1139/o83-049.
mRNAs for the two types of milk protein were determined in the polyadenylated (poly(A)+) and nonpolyadenylated (poly(A)-) RNA fractions prepared from lactating rat mammary gland, to assess if a high fraction of poly(A)- mRNA is a general characteristic of milk protein mRNAs. Use of the [3H]poly(U) assay to evaluate the effectiveness of oligo(dT)-cellulose and poly(U)-Sepharose for the preparation of these samples showed that similar, essentially poly(A)-free, RNA samples could be prepared with either resin when the appropriate sample to resin ratio was used. The poly(A) tracts in rat mammary poly(A)+ RNA displayed bimodal distribution with peaks at 40 and 80 nucleotides. Translation of these samples confirmed previous observations that mammary poly(A)-RNA contains high amounts of translational activity; however, it is shown that translation gives an overestimate of the mRNA activity in this fraction. Analyses with complementary DNAs indicated that only 24-35% of the casein mRNA and 14-33% of the alpha-lactalbumin mRNA are present in poly(A)-RNA. Milk protein mRNAs are, therefore, comparable with other abundant mRNAs. Evidence is presented indicating that partial degradation can greatly reduce the extent of polyadenylation with little effect on the amount and on the translational activity of the mRNA.
为了评估多聚腺苷酸化(poly(A)+)的mRNA是否是乳蛋白mRNA的普遍特征,我们测定了从泌乳大鼠乳腺制备的多聚腺苷酸化(poly(A)+)和非多聚腺苷酸化(poly(A)-)RNA组分中两种乳蛋白的mRNA。使用[3H]聚(U)分析法评估寡聚(dT)-纤维素和聚(U)-琼脂糖制备这些样品的效果,结果表明,当使用合适的样品与树脂比例时,两种树脂都能制备出相似的、基本不含poly(A)的RNA样品。大鼠乳腺poly(A)+ RNA中的多聚腺苷酸序列呈现双峰分布,峰值分别在40和80个核苷酸处。对这些样品的翻译证实了先前的观察结果,即乳腺poly(A)-RNA含有大量的翻译活性;然而,结果表明,翻译会高估该组分中的mRNA活性。用互补DNA进行的分析表明,只有24%-35%的酪蛋白mRNA和14%-33%的α-乳白蛋白mRNA存在于poly(A)-RNA中。因此,乳蛋白mRNA与其他丰富的mRNA相当。有证据表明,部分降解会大大降低多聚腺苷酸化的程度,而对mRNA的量和翻译活性影响很小。