Acheson N H
Proc Natl Acad Sci U S A. 1978 Oct;75(10):4754-8. doi: 10.1073/pnas.75.10.4754.
The bulk of late virus-specific RNA synthesized in polyoma virus-infected mouse cells is larger than a single strand of poloma DNA. The arrangement of viral nucleotide sequences in these giant polyoma RNAs was studied by electron microscopy of hybrids between purified high molecular weight viral RNA and the HindII-1 fragment of polyoma DNA, which contains 91% of the viral genome. Hybrid molecules containing a short single-stranded gap (corresponding to the 9% of viral sequences not present in HindII-1), flanked by double-stranded regions, were photographed and measured. The majority of hybrid molecules contained no single-stranded loops or branches, showing that all viral sequences are transcribed contiguously and that no nonviral sequences are present in the RNA. Hybrid molecules, containing RNA up to 3.5 times the genome length, had a repeating structure of single-stranded gaps 8% of genome length interspersed with double-stranded regions 89% of genome length, showing that giant polyoma RNAs contain tandem repeats of the nucleotide sequence of the entire viral DNA. A small proportion of hybrid molecules contained single-stranded branches or deletion loops in characteristic positions, indicating that RNA "splicing" may occur on high molecular weight nuclear polyoma RNA.
在多瘤病毒感染的小鼠细胞中合成的晚期病毒特异性RNA,其大部分比多瘤病毒DNA的单链要大。通过对纯化的高分子量病毒RNA与多瘤病毒DNA的HindII - 1片段(包含91%的病毒基因组)之间的杂交体进行电子显微镜观察,研究了这些巨大多瘤RNA中病毒核苷酸序列的排列。对含有短单链缺口(对应于HindII - 1中不存在的9%病毒序列)且两侧为双链区域的杂交分子进行拍照和测量。大多数杂交分子不包含单链环或分支,这表明所有病毒序列都是连续转录的,并且RNA中不存在非病毒序列。含有长达基因组长度3.5倍的RNA的杂交分子,具有单链缺口(占基因组长度的8%)与双链区域(占基因组长度的89%)相间的重复结构,这表明巨大多瘤RNA包含整个病毒DNA核苷酸序列的串联重复。一小部分杂交分子在特征位置含有单链分支或缺失环,这表明RNA“剪接”可能发生在高分子量核多瘤RNA上。