ANALYSIS OF REGIONS OF RAG-2 IMPORTANT FOR V(D)J RECOMBINATION
ANALYSIS OF REGIONS OF RAG-2 IMPORTANT FOR V(D)J RECOMBINATION
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DOI：
10.1093/nar/22.10.1810
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发表时间：
1994-05-25
影响因子：
14.9
通讯作者：
OETTINGER, MA
中科院分区：
文献类型：
--
作者：
CUOMO, CA;OETTINGER, MA
重组酶激活基因RAG-1和RAG-2共同作用以激活V（D）J重组，因此在免疫系统多样性的产生中发挥重要作用。作为理解RAG-2蛋白功能的第一步，我们已经测试了一系列缺失和插入突变诱导各种模型底物的V（D）J连接的能力。测定突变体诱导缺失和倒位V（D）J连接的能力，从而测试它们形成信号和编码接头的能力，并且在某些情况下，测试它们进行染色体外和整合重组底物的重组的能力。所有这些反应都受到任何一种突变的类似影响。虽然RAG-2蛋白在其长度上显示出广泛的进化保守性，但我们发现RAG-2的羧基末端部分，包括酸性区域，对于测试的所有形式的重组都是不稳定的。相反，我们在N-末端区域产生的所有突变都严重减少了重组。因此，V（D）J重组所需的核心活性区仅限于RAG-2蛋白的前四分之三。
The recombinase activating genes RAG-1 and RAG-2 operate together to activate V(D)J recombination, and thus play an essential role in the generation of immune system diversity. As a first step in understanding the function of the RAG-2 protein, we have tested a series of deletion and insertion mutations for their ability to induce V(D)J joining of a variety of model substrates. Mutants were assayed for their ability to induce deletional and inversional V(D)J joining, thereby testing their proficiency at forming both signal and coding joints, and, in some cases, for their ability to carry out recombination of both extrachromosomal and integrated recombination substrates. All these reactions were affected similarly by any one mutation. Although the RAG-2 protein shows extensive evolutionary conservation across its length, we found that the carboxy-terminal portion of RAG-2, including an acidic region, is dispensable for all forms of recombination tested. In contrast, all mutations we created in the N-terminal region severely decreased recombination. Thus, the core active region required for V(D)J recombination is confined to the first three-quarters of the RAG-2 protein.