Weis-Garcia F, Besmer E, Sawchuk D J, Yu W, Hu Y, Cassard S, Nussenzweig M C, Cortes P
Laboratory of Molecular Immunology, The Rockefeller University, New York, New York 10021, USA.
Mol Cell Biol. 1997 Nov;17(11):6379-85. doi: 10.1128/MCB.17.11.6379.
Antigen receptor genes are assembled through a mechanism known as V(D)J recombination, which involves two different joining reactions: signal and coding joining. Formation of these joints is essential for antigen receptor assembly as well as maintaining chromosomal integrity. Here we report on a cell-free system for coding joint formation using deletion and inversion recombination substrates. In vitro coding joint formation requires RAG1, RAG2, and heat-labile factors present in the nuclear extract of nonlymphoid cells. Both inversion- and deletion-mediated coding joint reactions produce diverse coding joints, with deletions and P nucleotide addition. We also show that deletion-mediated coding joint formation follows the 12/23 rule and requires the catalytic subunit of DNA-dependent protein kinase.
抗原受体基因通过一种称为V(D)J重组的机制进行组装，该机制涉及两种不同的连接反应：信号连接和编码连接。这些连接的形成对于抗原受体组装以及维持染色体完整性至关重要。在此，我们报道了一种使用缺失和倒位重组底物进行编码连接形成的无细胞系统。体外编码连接形成需要RAG1、RAG2以及非淋巴细胞核提取物中存在的热不稳定因子。倒位和缺失介导的编码连接反应都会产生多样的编码连接，伴有缺失和P核苷酸添加。我们还表明，缺失介导的编码连接形成遵循12/23规则，并且需要DNA依赖性蛋白激酶的催化亚基。
