An adenovirus type 5 gene function required for initiation of viral DNA replication.
An adenovirus type 5 gene function required for initiation of viral DNA replication.
复制标题
启动病毒 DNA 复制所需的 5 型腺病毒基因功能。
DOI：
10.1016/0042-6822(75)90443-2
复制
发表时间：
1975
期刊：
影响因子：
3.7
通讯作者：
J. Sussenbach
中科院分区：
文献类型：
--
作者：
P. C. Vliet;J. Sussenbach
用属于不同互补基团的DNA阴性温度敏感突变体(H5ts125和H5ts36)感染人和猴细胞后，研究了5型腺病毒(Ad5)的DNA复制。当在允许温度(32°)下进行感染，然后转换到非允许温度(39.5°)时，H5ts125感染细胞的病毒DNA合成在移位后1小时内下降90%，而H5ts36感染细胞的DNA合成仅在6小时后下降。对在抑制条件下合成的各种形式的DNA的分析表明，两个突变体的复制与成熟病毒DNA的比例都是恒定的，但没有观察到复制分子的积累。当H5ts125感染的细胞在32°或39.5°脉冲标记[~3H]胸腺嘧啶核苷，然后在39.5°追逐标记时，复制的DNA转化为成熟DNA的速度与野生型感染细胞相同。这表明链的传播和终止可以在不允许的条件下正常发生。在39.5°下进行的密度标记实验结果与H5ts125感染细胞在不允许的温度下的起始块一致。因此，H5ts125基因产物和H5ts36基因产物可能是启动新一轮复制所必需的。讨论了H5ts125基因编码的腺病毒特异性DNA结合蛋白的启动过程中的潜在作用。
Adenovirus type 5 (Ad5) DNA replication was studied after infection of human or monkey cells with two DNA-negative temperature-sensitive mutants belonging to different complementation groups (H5ts125 and H5ts36). When infection was carried out at the permissive temperature (32°) followed by a shift to the nonpermissive temperature (39.5°) viral DNA synthesis in H5ts125-infected cells was reduced 90% within 1 hr after shift-up, while a decline in DNA synthesis in H5ts36-infected cells is only observed after 6 hr. Analysis of the various forms of DNA synthesized under conditions of inhibition showed a constant ratio of replicating to mature viral DNA for both mutants, while no accumulation of replicating molecules was observed.When H5ts125-infected cells were pulse-labeled with [3H]thymidine at 32 or 39.5° followed by a chase of the label at 39.5°, replicating DNA was converted into mature DNA at the same rate as in wild-type-infected cells. This indicates that chain propagation and termination could occur normally under nonpermissive conditions. The results of density labeling experiments performed at 39.5° are in agreement with an initiation block in H5ts125-infected cells at the nonpermissive temperature. It is concluded that the H5ts125 gene product and possibly also the H5ts36 gene product are required for the initiation of new rounds of replication. The potential role in initiation of the adenovirus-specific DNA binding protein, which is coded for by the H5ts125 gene, is discussed.