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Publication : Distinct roles for RAG-1 in the initiation of V(D)J recombination and in the resolution of coding ends.

First Author  Gallo ML Year  1994
Journal  J Biol Chem Volume  269
Issue  35 Pages  22188-92
PubMed ID  8071342 Mgi Jnum  J:19967
Mgi Id  MGI:68086 Doi  10.1016/s0021-9258(17)31774-x
Citation  Gallo ML, et al. (1994) Distinct roles for RAG-1 in the initiation of V(D)J recombination and in the resolution of coding ends. J Biol Chem 269(35):22188-92
abstractText  Although RAG-1 and RAG-2 have been shown to be indispensible for V(D)J recombination, their exact role in this reaction remains unclear. Co-transfecting RAG-1 and RAG-2 expression vectors into NIH3T3 fibroblasts confers V(D)J recombination activity to these otherwise recombinationally inactive cells. In this report we have found that in transient transfections of mouse NIH3T3 fibroblasts with RAG-1 and RAG-2 and the appropriate recombination substrates, one RAG-1 expression vector, pRAG-1A, is capable of yielding both signal joints and coding joints, while another RAG-1 expression vector, pRAG-1B, yields only signal joints. The RAG-1 open reading frame for these two expression vectors is interchangeable, indicating that the inability to resolve coding joints is due to the 45-base pair difference found in the 5'-untranslated regions of these constructs. Differences in this region result in a 15-fold difference in gene expression when the luciferase coding region is substituted for the RAG-1 cDNA. This report provides evidence that RAG-1 may have a role in both the initiation of V(D)J recombination as well as the resolution of coding ends. The data also suggest that these RAG-1 activities may be dependent on different levels of RAG-1 expression.
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