Van Ness J, Maxwell I H, Hahn W E
Cell. 1979 Dec;18(4):1341-9. doi: 10.1016/0092-8674(79)90244-7.
The complexity of nonadenylated mRNA [poly(A)-mRNA] has been determined by hybridization with single-copy DNA (scDNA) and cDNA. Our results show that poly(A)- and poly(A)+ mRNA are essentially nonoverlapping (nonhomologous) sequence populations of similar complexity. The sum of the complexities of poly(A)+ mRNA and poly(A)- mRNA is equal to that of total polysomal RNA or total mRNA, or the equivalent of approximately 1.7 x 10(5) different sequences 1.5 kb in length. Poly(A)- mRNA, isolated from polysomal RNA by benzoylated cellulose chromatography, hybridized with 3.6% of the scDNA, corresponding to a complexity of 7.8 x 10(4) different 1.5 kb sequences. The equivalent of only one adenosine tract of approximately 20 nucleotides per 100 poly(A)- mRNA molecules 1.5 kb in size was observed by hybridization with poly(U). cDNA was transcribed from poly(A)- mRNA using random oligonucleotides as primers. Only 1-2% of the single-copy fraction of this cDNA was hybridized using poly(A)+ mRNA as a driver. These results show that poly(A)- mRNA shares few sequences with poly(A)+ mRNA and thus constitutes a separate, complex class of messenger RNA. These measurements preclude the presence of a complex class of bimorphic mRNAs [that is, species present in both poly(A)+ and poly(A)- forms] in brain polysomes.
通过与单拷贝DNA(scDNA)和cDNA杂交,已确定了非腺苷酸化mRNA([poly(A)-mRNA])的复杂性。我们的结果表明,poly(A)-和poly(A)+ mRNA本质上是具有相似复杂性的非重叠(非同源)序列群体。poly(A)+ mRNA和poly(A)- mRNA的复杂性总和等于总多核糖体RNA或总mRNA的复杂性,或相当于约1.7×10⁵个长度为1.5 kb的不同序列。通过苯甲酰化纤维素色谱法从多核糖体RNA中分离出的poly(A)- mRNA,与3.6%的scDNA杂交,相当于7.8×10⁴个不同的1.5 kb序列的复杂性。通过与poly(U)杂交观察到,每100个大小为1.5 kb的poly(A)- mRNA分子中,仅相当于一个约20个核苷酸的腺苷序列。使用随机寡核苷酸作为引物,从poly(A)- mRNA转录出cDNA。以poly(A)+ mRNA作为驱动子,该cDNA的单拷贝部分只有1 - 2%发生杂交。这些结果表明,poly(A)- mRNA与poly(A)+ mRNA共享的序列很少,因此构成了一类单独的、复杂的信使RNA。这些测量结果排除了脑多核糖体中存在一类复杂的双态mRNA(即同时以poly(A)+和poly(A)-形式存在的物种)的可能性。