Utilization of nonhomologous minus-strand DNA transfer to generate recombinant retroviruses.
Utilization of nonhomologous minus-strand DNA transfer to generate recombinant retroviruses.
复制标题
利用非同源负链 DNA 转移产生重组逆转录病毒。
DOI：
10.1128/jvi.71.3.2487-2494.1997
复制
发表时间：
1997
影响因子：
5.4
通讯作者：
Hu,WS
中科院分区：
文献类型：
--
作者：
Yin,PD;Pathak,VK;Rowan,AE;Teufel2nd,RJ;Hu,WS
在逆转录过程中，负链 DNA 转移连接位于病毒 RNA 两端的序列，产生长末端重复序列。人们认为位于病毒RNA序列两端的重复(R)区的同源性有利于负链DNA转移。在本报告中，研究了 R 区同源性降低对 DNA 合成和病毒滴度的影响。构建了逆转录病毒载体 PY31，其中包含来自莫洛尼鼠肉瘤病毒和脾坏死病毒的 5' 和 3' 顺式作用元件。这两种病毒在遗传上是不同的，并且两个 R 区几乎没有同源性。在一轮复制中，PY31 滴度比具有高度同源性 R 区的对照载体低约 3,000 倍。对未整合的 DNA 和整合的原病毒中负链 DNA 转移连接点的分子特征进行了分析。在转移连接处发现了短段同源性，并且可能用于促进负链 DNA 转移。观察到负链强终止DNA和弱终止DNA均介导链转移。 PY31完成逆转录的能力表明负链DNA转移可用于连接来自两种不同病毒的序列以形成重组病毒。这些结果表明基因不同的病毒可以通过这种机制相互作用的可能性。
During reverse transcription, minus-strand DNA transfer connects the sequences located at the two ends of the viral RNA to generate a long terminal repeat. It is thought that the homology in the repeat (R) regions located at the two ends of the viral RNA sequences facilitate minus-strand DNA transfer. In this report, the effects of diminished R-region homology on DNA synthesis and virus titer were examined. A retrovirus vector, PY31, was constructed to contain the 5' and 3' cis-acting elements from Moloney murine sarcoma virus and spleen necrosis virus. These two viruses are genetically distinct, and the two R regions contain little homology. In one round of replication, the PY31 titer was approximately 3,000-fold lower than that of a control vector with highly homologous R regions. The molecular characteristics of the junctions of minus-strand DNA transfer were analyzed in both unintegrated DNA and integrated proviruses. Short stretches of homology were found at the transfer junctions and were likely to be used to facilitate minus-strand DNA transfer. Both minus-strand strong-stop DNA and weak-stop DNA were observed to mediate strand transfer. The ability of PY31 to complete reverse transcription indicates that minus-strand DNA transfer can be used to join sequences from two different viruses to form recombinant viruses. These results suggest the provocative possibility that genetically distinct viruses can interact through this mechanism.