Effects of limiting homology at the site of intermolecular recombinogenic template switching during moloney murine leukemia virus replication
Effects of limiting homology at the site of intermolecular recombinogenic template switching during moloney murine leukemia virus replication
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DOI：
10.1128/jvi.75.23.11263-11274.2001
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发表时间：
2001-12-01
影响因子：
5.4
通讯作者：
Telesnitsky, A
中科院分区：
文献类型：
--
作者：
Pfeiffer, JK;Telesnitsky, A
建立了基于Moloney小鼠白血病病毒的单复制周期实验，研究了限制模板和引物链互补程度对重组模板切换的影响。该系统模拟了强制复制选择重组，在这种重组中，新生DNA从供体模板的末端转移到另一个共包装RNA上的受体位置。当测试与转移DNA具有不同程度互补的受体靶区时，有效的重组发生在14个互补核苷酸上。确定了正确靶向、转移相关错误、错配扩展和在到达供体模板末端之前转移的频率。所有四个分子事件都发生了，它们的比例取决于受体/转移DNA互补性的性质。当互补性严重受限时，重组效率低下，大多数产物是由异常的第二链转移引起的，而不是rna之间的强制模板切换。当受体和供体之间的同源性有限时，还观察到其他类型的逆转录产物，包括病毒和宿主序列之间模板转换产生的一些产物。
A Moloney murine leukemia virus-based single-replication-cycle assay was developed to study the effects of limiting the extent of template and primer strand complementarity on recombinogenic template switching. This system mimicked forced copy choice recombination in which nascent DNA transfers from the end of a donor template to an acceptor position on the other copackaged RNA. When acceptor target regions with different extents of complementarity to the transferring DNA were tested, efficient recombination occurred with as few as 14 complementary nucleotides. The frequencies of correct targeting, transfer-associated errors, mismatch extension, and transfer before reaching the end of the donor template were determined. All four molecular events occurred, with their proportions varying depending on the nature of acceptor/transferring DNA complementarity. When complementarity was severely limited, recombination was inefficient and most products resulted from aberrant second-strand transfer rather than from forced template switching between RNAs. Other classes of reverse transcription products, including some that resulted from template switching between virus and host sequences, were also observed when homology between the acceptor and donor was limited.