Fugmann S D, Lee A I, Shockett P E, Villey I J, Schatz D G
Howard Hughes Medical Institute, Section of Immunobiology, Yale University School of Medicine, New Haven, Connecticut 06520-8011, USA.
Annu Rev Immunol. 2000;18:495-527. doi: 10.1146/annurev.immunol.18.1.495.
V(D)J recombination proceeds through a series of protein:DNA complexes mediated in part by the RAG1 and RAG2 proteins. These proteins are responsible for sequence-specific DNA recognition and DNA cleavage, and they appear to perform multiple postcleavage roles in the reaction as well. Here we review the interaction of the RAG proteins with DNA, the chemistry of the cleavage reaction, and the higher order complexes in which these events take place. We also discuss postcleavage functions of the RAG proteins, including recent evidence indicating that they initiate the process of coding end processing by nicking hairpin DNA termini. Finally, we discuss the evolutionary and functional implications of the finding that RAG1 and RAG2 constitute a transposase, and we consider RAG protein biochemistry in the context of several bacterial transposition systems. This suggests a model of the RAG protein active site in which two divalent metal ions serve alternating and opposite roles as activators of attacking hydroxyl groups and stabilizers of oxyanion leaving groups.
V(D)J 重排通过一系列蛋白质:DNA 复合物进行,部分由 RAG1 和 RAG2 蛋白介导。这些蛋白质负责序列特异性 DNA 识别和 DNA 切割,并且它们似乎在反应中也执行多种切割后功能。在这里,我们综述了 RAG 蛋白与 DNA 的相互作用、切割反应的化学过程以及这些事件发生的高阶复合物。我们还讨论了 RAG 蛋白的切割后功能,包括最近的证据表明它们通过切割发夹 DNA 末端启动编码末端加工过程。最后,我们讨论了 RAG1 和 RAG2 构成转座酶这一发现的进化和功能意义,并在几种细菌转座系统的背景下考虑 RAG 蛋白生物化学。这提出了一个 RAG 蛋白活性位点模型,其中两个二价金属离子作为攻击羟基的激活剂和氧阴离子离去基团的稳定剂发挥交替且相反的作用。