Canaani D, Kahana C, Mukamel A, Groner Y
Proc Natl Acad Sci U S A. 1979 Jul;76(7):3078-82. doi: 10.1073/pnas.76.7.3078.
The 5'-cap-containing leader sequence of the most abundant 19S and 16S mRNAs of simian virus 40 (SV40) was previously mapped between 0.67 and 0.76 map units. We now find that the two late mRNA species contain multiple 5' ends. Eight different RNase T2-resistant cap structures were identified:m7GpppmAmpU (47%); m7GpppmAmpUmpU (19%); m7GpppmAmpC (16%); m7GpppmAmpCmpA (5%); m7GpppmAmpG (6%); m7GpppGmpC (3%); m7GpppmAmGmpA (2%); m7GpppGmpCmpG (2%). Capped T1 oligonucleotides of 19S and 16S mRNAs have been isolated by two different procedures: (i) chromatography on a DEAE-cellulose column followed by paper electrophoresis and (ii) two-dimensional electrophoresis/homochromatography. Cap structures of the isolated 5' oligonucleotides were identified. Each of the major caps was found to be associated with a few differential 5' oligonucleotides, implying a vast heterogeneity at the termini of SV40 late mRNAs. The results suggest that on SV40 DNA, RNA polymerase II has a reportoire of initiation points. In most of the cases, initiation takes place with adenosine triphosphate followed by a pyrimidine. Alternatively, transcription may start at one specific point but a unique mechanism of processing generates heterogeneous populations of termini with a common 5' adenosine triphosphate.
猴病毒40(SV40)最丰富的19S和16S mRNA含5'-帽的前导序列先前被定位在0.67至0.76个图距单位之间。我们现在发现这两种晚期mRNA种类含有多个5'末端。鉴定出了八种不同的抗核糖核酸酶T2帽结构:m7GpppmAmpU(47%);m7GpppmAmpUmpU(19%);m7GpppmAmpC(16%);m7GpppmAmpCmpA(5%);m7GpppmAmpG(6%);m7GpppGmpC(3%);m7GpppmAmGmpA(2%);m7GpppGmpCmpG(2%)。19S和16S mRNA的带帽T1寡核苷酸已通过两种不同方法分离:(i)在二乙氨基乙基纤维素柱上进行色谱分离,随后进行纸电泳;(ii)二维电泳/同系色谱法。鉴定了分离出的5'寡核苷酸的帽结构。发现每个主要帽都与一些不同的5'寡核苷酸相关联,这意味着SV40晚期mRNA末端存在巨大的异质性。结果表明,在SV40 DNA上,RNA聚合酶II有一系列起始点。在大多数情况下,起始是由三磷酸腺苷接着一个嘧啶发生的。或者,转录可能从一个特定点开始,但一种独特的加工机制产生了具有共同5'三磷酸腺苷的异质末端群体。