Cue D, Feiss M
Department of Microbiology, University of Iowa, Iowa City, IA 52242, USA.
J Mol Biol. 1998 Jul 3;280(1):11-29. doi: 10.1006/jmbi.1998.1841.
Termination of packaging of the lambda chromosome involves completion of translocation of the DNA into the head shell, and conversion of the translocation complex into a cleavage complex. The cleavage reaction introduces staggered nicks into the downstream cosN to generate the right cohesive end of the chromosome. cosQ, a site adjacent to cosN, was found to be required for nicking the bottom strand of cosN; bottom strand nicking was also sequence-specific for bps at the nick site. Nicking of the top strand of cosN (cosNL) was stimulated by cosQ, but fidelity and efficiency of cosNL nicking were largely dictated by other cos subsites (i.e. cosB and I2). Aberrant top-strand cleavage within cosQ was observed in the absence of I2, and nicking at a site 8 nt 5' to the normal cosNL nick site occurred in the absence of cosB. The presence of cosQ was found to be insufficient to arrest DNA translocation in vivo, indicating that cosQ, per se, is not a packaging stop signal. A model is presented in which the role of cosQ is to depolarize the asymmetric arrangement of terminase protomers in the translocation complex so that protomers are configured to match the 2-fold rotational symmetry of cosN.
λ染色体包装的终止涉及DNA向头部外壳的转位完成,以及转位复合物向切割复合物的转变。切割反应在下游cosN处引入交错切口,以产生染色体的右粘性末端。发现与cosN相邻的位点cosQ是切割cosN底部链所必需的;底部链切割对切口位点的碱基对也具有序列特异性。cosQ刺激cosN顶部链(cosNL)的切割,但cosNL切割的保真度和效率在很大程度上由其他cos亚位点(即cosB和I2)决定。在没有I2的情况下,观察到cosQ内的异常顶部链切割,在没有cosB的情况下,在正常cosNL切口位点5'端8个核苷酸处发生切割。发现cosQ的存在不足以在体内阻止DNA转位,这表明cosQ本身不是一个包装终止信号。本文提出了一个模型,其中cosQ的作用是使转位复合物中末端酶原的不对称排列去极化,从而使原聚体配置成与cosN的2倍旋转对称性相匹配。