A basic motif in the N-terminal region of RAG1 enhances V(D)J recombination activity

被引:55
作者
McMahan, CJ
Difilippantonio, MJ
Rao, N
Spanopoulou, E
Schatz, DG
机构
[1] YALE UNIV,SCH MED,HOWARD HUGHES MED INST,NEW HAVEN,CT 06520
[2] YALE UNIV,SCH MED,IMMUNOBIOL SECT,NEW HAVEN,CT 06520
[3] YALE UNIV,SCH MED,DEPT GENET,NEW HAVEN,CT 06520
[4] CUNY MT SINAI SCH MED,RUTTENBERG CANC CTR,NEW YORK,NY 10029
关键词
D O I
10.1128/MCB.17.8.4544
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The variable portions of antigen receptor genes are assembled from component gene segments by a site-specific recombination reaction known as V(D)J recombination. The RAG1 and RAG2 proteins are the critical lymphoid cell-specific components of the recombination enzymatic machinery and are responsible for site-specific DNA recognition and cleavage. Previous studies had defined a minimal, recombinationally active core region of murine RAG1 consisting of amino acids 384 to 1008 of the 1,040-residue RAG1 protein. No recombination function has heretofore been ascribed to any portion of the 383-amino-acid N-terminal region that is missing from the core, but it seems likely to be of functional significance, based on its evolutionary conservation. Using extrachromosomal recombination substrates, we demonstrate here that the N-terminal region enhances the recombination activity of RAG1 by up to an order of magnitude in a variety of cell lines, Deletion analysis localized a region of the N terminus critical for this effect to amino acids 216 to 238, and further mutagenesis demonstrated that a small basic amino acid motif (BIIa) in this region is essential for enhancing the activity of RAG1. Despite the fact that BIIa is important for the interaction of RAG1 with the nuclear localization factor Srp-1, it does not appear to enhance recombination by facilitating nuclear transport of RAG1. A variety of models for how this region stimulates the recombination activity of RAG1 are considered.
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页码:4544 / 4552
页数:9
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