Takano T, Hatanaka M
Proc Natl Acad Sci U S A. 1975 Jan;72(1):343-7. doi: 10.1073/pnas.72.1.343.
[3H]Uridine-labeled Rauscher leukemia virus was used to infect mouse embryo fibroblasts. After the infected cells were separated into nuclear and cytoplasmic fractions nucleic acid was extracted by sodium dodecyl sulfate-phenol-chloroform treatment and analyzed by Cs2SO4 and sucrose density gradient centrifugation. Between 45 and 70 min after infection a transient and synchronized shift of the acid-insoluble radioactive peak toward the RNA-DNA hybrid region occurred in both the nuclear and cytoplasmic fractions. The density of the cytoplasmic hybrid shifted to 1.56 g/ml (RNA equals about 50%), while the sedimentation rate decreased from 36 S to 14 S; however, the density of the nuclear hybrid shifted to 1.58-1.48 g/ml (RNA equals 57-17%, respectively), while its sedimentation rate remained about 65 S. The hybrids in both the nuclear and the cytoplasmic fractions still showed hybrid density after heat denaturation. The processes of the early stages of RNA tumor virus infection are discussed with regard to the functions of viral RNA-dependent DNA polymerase (reverse transcriptase) and a possible integration of viral genetic information into the host chromosome.
用[3H]尿苷标记的劳氏肉瘤病毒感染小鼠胚胎成纤维细胞。感染细胞分离成细胞核和细胞质部分后,通过十二烷基硫酸钠-苯酚-氯仿处理提取核酸,并通过硫酸铯和蔗糖密度梯度离心进行分析。感染后45至70分钟之间,细胞核和细胞质部分中酸不溶性放射性峰向RNA-DNA杂交区域发生短暂且同步的转移。细胞质杂交体的密度转移至1.56克/毫升(RNA约占50%),而沉降率从36S降至14S;然而,细胞核杂交体的密度转移至1.58 - 1.48克/毫升(RNA分别占57% - 17%),而其沉降率仍约为65S。热变性后,细胞核和细胞质部分中的杂交体仍显示杂交密度。关于病毒RNA依赖性DNA聚合酶(逆转录酶)的功能以及病毒遗传信息可能整合到宿主染色体中的情况,对RNA肿瘤病毒感染早期阶段的过程进行了讨论。