Hagemann A T, Rosenberg S M
Institute of Molecular Biology, University of Oregon, Eugene 97403.
Genetics. 1991 Nov;129(3):611-21. doi: 10.1093/genetics/129.3.611.
Heteroduplex patch recombinants have received information in one DNA chain but have not recombined flanking markers. Evidence regarding which chain is exchanged bears on the structure of recombination intermediates. The direction of travel along DNA of RecBCD recombinase, the central enzyme in the Escherichia coli RecBCD pathway of homologous recombination, is determined in phage lambda by the orientation of the packaging origin, cos. cos is a double-chain cut site which serves as a preferred entry site for RecBCD. Using partially denaturing gels to resolve heteroduplex molecules, we have examined patch recombinants at the lambda ren gene. We report that the transferred information in Chi-stimulated patches at ren can occur on either chain, but is biased to the chain ending 5' at the right of the lambda map (the lambda r chain) in phage carrying cos in its normal orientation. The chain bias switches in favor of the chain that ends 3' at the right (the lambda l chain) when RecBCD travel direction is reversed by inverting cos. We entertain models that accommodate these and other results pertaining to the structure of RecBCD-mediated recombinants.
异源双链修补重组体在一条DNA链中获得了信息,但未重组侧翼标记。关于哪条链发生交换的证据与重组中间体的结构有关。沿着RecBCD重组酶的DNA行进方向,RecBCD重组酶是大肠杆菌同源重组RecBCD途径中的核心酶,在噬菌体λ中由包装起点cos的方向决定。cos是一个双链切割位点,作为RecBCD的首选进入位点。使用部分变性凝胶来解析异源双链分子,我们在λ ren基因处检查了修补重组体。我们报告说,在ren处受Chi刺激的修补中转移的信息可以出现在任何一条链上,但在正常方向携带cos的噬菌体中,偏向于在λ图谱右侧5'端结束的链(λ r链)。当通过反转cos来逆转RecBCD的行进方向时,链偏向会转而有利于在右侧3'端结束的链(λ l链)。我们考虑了一些模型,这些模型能够解释这些以及其他与RecBCD介导的重组体结构相关的结果。