Dina D, Beemon K, Duesberg P
Cell. 1976 Oct;9(2):299-309. doi: 10.1016/0092-8674(76)90120-3.
The 50S-70S RNA of a Moloney sarcoma-leukemia virus [Mo-MSV(MLV)] complex produced by a particular mouse cell line was shown by gel electrophoresis to contain a major (97%) 30S sarcoma-specific subunit species and a minor (3%) 38S leukemia virus-specific subunit. On the basis of its sedimentation coefficient and known complexity, the 30S Mo-MSV RNA was estimated to be a unique RNA molecule of about 6000 nucleotides. Hybridization experiments using viral RNA and DNA complementary to viral RNA (cDNA) made by viral DNA polymerase indicated that the 30S Mo-MSV RNA shared 70% of its sequences with Mo-MLV, 30% with another MLV derived from Mo-MLV, and 30% with Kirsten sarcoma-xenotropic leukemia virus. The 30S Mo-MSV RNA sequences shared with these viruses were not additive. The Tm of a Mo-MSV RNA-MLV cDNA hybrid was 83 degrees C, indicating that large contiguous nucleotide sequences were shared between the two nucleic acids. Mo-MSV RNA and Mo-MLV RNA shared possibly seven of 20-30 RNAase T1-resistant oligonucleotides, while Mo-MSV RNA contained three, and Mo-MLV RNA contained at least five specific oligonucleotides. We conclude that the 30S Mo-MSV RNA molecule consists of approximately 70% (about 4200 nucleotides) Mo-MLV-specific sequences and of 30% (1800 nucleotides) Mo-MSV-specific sequences covalently linked. Our results favor the hypothesis that 30S Mo-MSV RNA was generated by recombination between Mo-MLV and other genetic elements. We discuss whether all or only the MSV-specific sequences of the 30S Mo-MSV RNA function as sarcoma genes. Mo-MLV cDNA was hybridized about 45% by unfractionated Mo-MSV (MLV) RNA at RNA/DNA ratios of up to 10, about 50% by electrophoretically purified 30S Mo-MSV RNA at RNA/DNA ratios up to 500, but close to 100% by unfractionated Mo-MSV(MLV) RNA at RNA/DNA ratios over 900. This indicated that unfractionated RNA of our Mo-MSV(MLV) contained a complete complement of Mo-MLV, albeit at a low ratio.
通过凝胶电泳显示,由特定小鼠细胞系产生的莫洛尼肉瘤 - 白血病病毒[Mo - MSV(MLV)]复合物的50S - 70S RNA含有一个主要的(97%)30S肉瘤特异性亚基种类和一个次要的(3%)38S白血病病毒特异性亚基。根据其沉降系数和已知复杂性,估计30S Mo - MSV RNA是一个约6000个核苷酸的独特RNA分子。使用病毒RNA和由病毒DNA聚合酶产生的与病毒RNA互补的DNA(cDNA)进行的杂交实验表明,30S Mo - MSV RNA与Mo - MLV共享70%的序列,与另一种源自Mo - MLV的MLV共享30%的序列,与柯斯顿肉瘤 - 嗜异性白血病病毒共享30%的序列。30S Mo - MSV RNA与这些病毒共享的序列不是累加的。Mo - MSV RNA - MLV cDNA杂交体的熔解温度为83℃,表明两种核酸之间共享大的连续核苷酸序列。Mo - MSV RNA和Mo - MLV RNA可能共享20 - 30个对核糖核酸酶T1有抗性的寡核苷酸中的7个,而Mo - MSV RNA含有3个,Mo - MLV RNA含有至少5个特异性寡核苷酸。我们得出结论,30S Mo - MSV RNA分子由大约70%(约4200个核苷酸)的Mo - MLV特异性序列和30%(1800个核苷酸)的Mo - MSV特异性序列共价连接而成。我们的结果支持这样的假设,即30S Mo - MSV RNA是由Mo - MLV与其他遗传元件之间的重组产生的。我们讨论了30S Mo - MSV RNA的所有序列还是仅MSV特异性序列作为肉瘤基因起作用。在RNA/DNA比例高达10时,未分级的Mo - MSV(MLV) RNA与Mo - MLV cDNA的杂交率约为45%,在RNA/DNA比例高达500时,经电泳纯化的30S Mo - MSV RNA与Mo - MLV cDNA的杂交率约为50%,但在RNA/DNA比例超过900时,未分级的Mo - MSV(MLV) RNA与Mo - MLV cDNA的杂交率接近100%。这表明我们的Mo - MSV(MLV)未分级RNA含有完整的Mo - MLV互补序列,尽管比例较低。