Integration of avian sarcoma virus DNA in chicken cells.
Integration of avian sarcoma virus DNA in chicken cells.
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禽肉瘤病毒 DNA 在鸡细胞中的整合。
DOI：
10.1016/0042-6822(81)90539-0
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发表时间：
1981
期刊：
影响因子：
3.7
通讯作者：
Varmus,HE
中科院分区：
文献类型：
--
作者：
Hughes,SH;Vogt,PK;Stubblefield,E;Bishop,JM;Varmus,HE
我们使用针对禽肉瘤病毒（ASV）基因组各个区域的限制性核酸内切酶和杂交试剂来评估在培养物感染后在鸡细胞（该病毒的天然宿主）中发现的原病毒的结构和整合位点。 ASV 转化的鸡成纤维细胞集落显示出在宿主基因组的一到几个位点上含有新的原病毒；在所研究的九个菌落中，每个菌落的整合位点似乎有所不同。从大量培养中感染 ASV 的类淋巴母细胞中分离染色体也未能揭示 ASV 原病毒插入的首选位点。因此，在鸡细胞以及异源细胞中[Hughes S. H. et al.(1978).Cell15, 1397–1410;Sabran, J. L.,et al.(1979).J. Virol.29, 170–185;Martin, S.et al.(1979).Virology,96, 530–546;Gilmer, T., 和 Parsons, J. T. (1979).J. Virol.32, 762–770]，细胞 DNA 的许多区域可以容纳 ASV 原病毒，并且这些区域分布在多个染色体上。尽管在细胞 DNA 的许多位点都发现了原病毒，但在受感染的鸡细胞中，原病毒 DNA 的结构似乎是不变的。病毒 DNA 与大约末端或末端附近的细胞 DNA 连接。 330 个核苷酸的序列在未整合的线性 DNA 末端重复。重复结构由源自病毒RNA 3'和5'端的序列组成。 ASV DNA 的最终结构可以写成“cell DNA-3'5'-gag-pol-env-src-3'5'-cell DNA”，这与在不允许的异源宿主细胞中对 ASV 原病毒的研究类似地推断出[Hughes, S. H.,et al.(1978).Cell15, 1397–1410]。感染后3至6天，通过将细胞悬浮在琼脂中获得详细检查的集落。根据整合原病毒的物理图谱判断，这些菌落中的一些但不是全部似乎是克隆的。根据此标准，从感染后直接置于琼脂中的细胞获得的集落并未出现克隆；考虑对这种异常行为的解释。
We have used restriction endonucleases and hybridization reagents specific for various regions of the avian sarcoma virus (ASV) genome to assess the structure and sites of integration of proviruses found in chicken cells, the natural host for thiss viris, after infection in culture. Colonies of ASV-transformed chicken fibroblasts were shown to contain new proviruses located at from one to several sites in the host genome; the sites of integration appeared to differ in each of the nine colonies studied. Fractionation of chromosomes from lymphoblastoid cells infected in mass culture with ASV also failed to reveal preferred sites for insertion of ASV proviruses. Thus, in chicken cells, as well as in heterologous cells[Hughes S. H.et al.(1978).Cell15, 1397–1410;Sabran, J. L.,et al.(1979).J. Virol.29, 170–185;Martin, S.et al.(1979).Virology,96, 530–546;Gilmer, T., and Parsons, J. T. (1979).J. Virol.32, 762–770], many regions of cellular DNA can accomodate an ASV provirus, and these regions are distributed over several chromosomes. Although proviruses are found at many sites in cellular DNA, the structure of proviral DNA appears to be invariant in infected chicken cells. Viral DNA is linked to cellular DNA near or at the extremities of a ca. 330-nucleotide sequence repeated at the ends of unintegrated linear DNA. The repeated structure is composed of sequences derived from both the 3′ and the 5′ ends of viral RNa. The resulting structure for ASV DNA can be written “cell DNA-3′5′-gag-pol-env-src-3′5′-cell DNA,” as similarly deduced from studies of ASV proviruses in nonpermissive, heterologous host cells[Hughes, S. H.,et al.(1978).Cell15, 1397–1410]. The colonies examined in detail were obtained by suspending cells in agar 3 to 6 days after infection. Some but not all of these colonies appeared to be clonal, as judged by the physical mapping of integrated proviruses. Colonies obtained from cells placed in agar directly after infection did not appear clonal by this criterion; explanations for this anomalous behavior are considered.