SITE-SPECIFIC RECOMBINATION INTERMEDIATES TRAPPED WITH SUICIDE SUBSTRATES
SITE-SPECIFIC RECOMBINATION INTERMEDIATES TRAPPED WITH SUICIDE SUBSTRATES
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DOI：
10.1016/0092-8674(87)90336-9
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发表时间：
1987-08-28
期刊：
影响因子：
64.5
通讯作者：
LANDY, A
中科院分区：
文献类型：
--
作者：
NUNESDUBY, SE;MATSUMOTO, L;LANDY, A
设计了一系列新型底物，使中间体在位点特异性重组中有效积累。策略性放置的切口允许这些“自杀亚态”启动反应，但阻止其完成或逆转。因此，可以确定.lambda。位点特异性重组通过一对顺序的单链交换进行。这些结果排除了需要对所有四条DNA链进行协调切割的一类模型。序列链交换按照严格规定的顺序进行，这在整合重组和分离重组中都是相同的。这种特定的顺序似乎是由链交换位点远端的结合蛋白的排列所控制的。此外，当提供适当的5 " OH受体时，整合酶蛋白具有在非互反反应中执行单链DNA转移的能力。
A family of novel substrates was designed to enable the efficent accumulation of intermediates in site-specific recombination. Strategically placed nicks allow these "sucide substates" to initate the reaction but prevent its completion or reversal. Consequently, it has been possible to determine that .lambda. site-specific recombination proceeds by a pair of sequential single-stranded exchanges. These results rule out that class of models invoking a concerted cutting of all four DNA strands. The sequential strand exchanges are executed in a strictly prescribed order that is the same in both integrative and excisive recombination. This specified order appears to be governed by the arrangement of bound proteins distal to the sites of strands exchange. Furthermore, when provided with an appropriate 5'' OH acceptor, the Integrase protein has the capacity to executive a single DNA strand transfer in a nonreciprocal reaction.