Herpesvirus-dependent amplification and inversion of cell-associated viral thymidine kinase gene flanked by viral a sequences and linked to an origin of viral DNA replication.
Herpesvirus-dependent amplification and inversion of cell-associated viral thymidine kinase gene flanked by viral a sequences and linked to an origin of viral DNA replication.
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细胞相关病毒胸苷激酶基因的疱疹病毒依赖性扩增和倒位，其两侧为病毒 a 序列，并与病毒 DNA 复制起点相连。
DOI：
10.1073/pnas.79.18.5626
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发表时间：
1982
影响因子：
11.1
通讯作者：
Roizman,B
中科院分区：
文献类型：
--
作者：
Mocarski,ES;Roizman,B
单纯疱疹病毒1型或2型的基因组由两个组分L和S组成，它们在感染期间相对于彼此反转。因此，病毒DNA由四个等摩尔的分子群体组成，其不同之处仅在于L和S组分的相对取向。已有的研究表明，位于基因组末端的相同方向和位于L-S连接处的反向方向的a序列在L和S组分的反转中起关键作用。在这份报告中，我们描述了一个病毒依赖的系统，旨在允许识别的病毒基因能够反式反转DNA侧翼的序列的反向拷贝。在该系统中，用嵌合质粒将细胞转化为胸苷激酶阳性表型，所述嵌合质粒携带胸苷激酶基因，所述胸苷激酶基因侧翼为α序列的反向拷贝，并连接至源自S组分的病毒DNA复制起点。引入细胞的DNA以其原始序列排列作为头-尾多联体保留和繁殖。用单纯疱疹病毒1型或2型感染这些细胞，导致质粒序列扩增多达100倍，并使两侧有α序列拷贝的DNA倒位。在受感染的细胞中，扩增的常驻DNA以头-尾多联体的形式积累，并且除了倒位之外没有检测到重排。这些结果表明，依赖于序列的倒位需要反式作用的病毒基因产物。
The genome of herpes simplex virus 1 or 2 consists of two components, L and S, which invert relative to each other during infection. As a result, viral DNA consists of four equimolar populations of molecules differing solely in the relative orientations of the L and S components. Previous studies have shown that the a sequences, located in the same orientation at the genomic termini and in inverted orientation at the L-S junction, play a key role in the inversion of L and S components. In this report we describe a virus-dependent system designed to allow identification of the viral genes capable of acting in trans to invert DNA flanked by inverted copies of a sequences. In this system, cells are converted to the thymidine kinase-positive phenotype with a chimeric plasmid carrying the thymidine kinase gene flanked by inverted copies of the a sequence and linked to an origin of viral DNA replication derived from the S component. The DNA introduced into the cells is retained and propagated in its original sequence arrangement as head-to-tail concatemers. Infection of these cells with herpes simplex virus 1 or 2 results in as much as 100-fold amplification of the plasmid sequences and inversion of the DNA flanked by copies of the a sequence. In infected cells, the amplified resident DNA accumulates in head-to-tail concatemers and no rearrangement other than the inversions could be detected. These results suggest that the a sequence-dependent inversions required trans-acting viral gene products.