RAG1 and RAG2 cooperate in specific binding to the recombination signal sequence in vitro
RAG1 and RAG2 cooperate in specific binding to the recombination signal sequence in vitro
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DOI：
10.1074/jbc.274.11.7025
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发表时间：
1999-03-12
影响因子：
4.8
通讯作者：
Sadofsky, MJ
中科院分区：
文献类型：
--
作者：
Mo, XM;Bailin, T;Sadofsky, MJ
脊椎动物免疫系统发育的一个重要步骤是抗原受体基因的DNA水平重排。这一过程被称为“V（D）J重组”，在适当的位点由两种蛋白质RAG 1和RAG 2介导DNA切割。我们在这里报告说，这两种蛋白质合作，结合DNA显着更高的特异性比单独的蛋白质。在不存在任何交联剂的情况下进行三重复合物的凝胶纯化。这两种蛋白质仍然存在于复合物中，并且使用含碘尿苷的探针进行的UV交联显示RAG 1在七聚体和九聚体基序中都进行了密切接触，这两种蛋白质也显示出在没有任何DNA的情况下彼此缔合。这些发现完善了我们对伴随重组信号切割的蛋白质-DNA相互作用的理解。
An essential step in the development of the vertebrate immune system is the DNA level rearrangement of the antigen receptor genes. This process, termed "V(D)J recombination," beans with DNA cleavage at the appropriate sites mediated by the two proteins RAG1 and RAG2. We report here that the two proteins cooperate to bind DNA with significantly higher specificity than either protein alone. Gel purification of the triple complex is performed in the absence of any cross-linking agents. Both proteins remain present in the complex, and UV cross-linking using iodouridine-containing probes shows that RAG1 makes close contacts in both the heptamer and nonamer motifs, The two proteins are also shown to associate with each other in the absence of any DNA. These findings refine our understanding of the protein-DNA interactions that accompany cleavage at the recombination signals.