蛋白质相互作用在大肠杆菌的基因重组。交互涉及侦察和RecR克服RecA通过单链dna结合蛋白的抑制。
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Umezu K, Kolodner RD
蛋白质相互作用在大肠杆菌的基因重组。交互涉及侦察和RecR克服RecA通过单链dna结合蛋白的抑制。
J生物化学杂志。1994年11月25日,269(47):30005 - 13所示。
- PubMed ID
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7962001 (在PubMed]
- 文摘
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RecA促进同源配对的单链DNA (ssDNA)与双链DNA (dsDNA)。这个反应发生效率低下,如果ssDNA衬底与大肠杆菌preincubated ssDNA-binding蛋白质(单边带)。然而,侦察和RecR能为RecA作为辅助因素共同行动来克服这种抑制的单边带(Umezu K。、气、N.-W。Kolodner, r . d . (1993) Proc。国家的。学会科学。美国90年,3875 - 3879)。阐明这个过程的机制,我们研究蛋白质之间的相互作用RecA, RecF,侦察,RecR,单边带,ssDNA复合物的结构和活动特征与这些蛋白质的不同组合形成。我们得到以下结果。(我)重建身体与RecR和单边带。侦察和单边带比之间的交互RecO-RecR交互。 (ii) RecO and RecR do not remove SSB from SSB.ssDNA complexes, but instead bind to these complexes. The resulting RecO.RecR.SSB.ssDNA complexes were more active in RecA-mediated joint molecule formation than were SSB.ssDNA complexes. (iii) RecA can nucleate on the RecO.RecR.SSB.ssDNA complexes more efficiently than on SSB.ssDNA complexes. (iv) When RecA presynaptic filaments were formed in the presence of SSB, RecO, and RecR, the protein-DNA complexes obtained contained 70% of the amount of RecA required to saturate ssDNA. These complexes, however, can mediate joint molecule formation and strand exchange as efficiently as presynaptic filaments which are fully saturated with RecA. Based on these results, we propose dual roles for RecO and RecR in joint molecule formation. First, RecO and RecR bind to SSB.ssDNA complexes and modify their structure to allow RecA to nucleate on them efficiently. Second, RecO and RecR are retained in RecA presynaptic filaments and play a role in the subsequent homologous pairing process promoted by RecA.