The DNA-binding ability of HIVEP3/KRC decreases upon activation of V(D)J recombination

Somatic V(D)J recombination of the immune receptor genes is mediated by the recombination signal sequence (RSS) and the recombination-activating genes RAG1 and RAG2. Previously, proteins binding specifically to the RSS have been characterized in nuclear extracts of T and B lymphocytes. Further elucidation of the role of those RSS-binding proteins in V(D)J recombination, however, has been hampered by the fact that their identities have not been established. Here, we show that the major RSS-binding protein present in the nuclear extracts of B lymphocytes is an M_r 135,000 species. Notably, its affinity for the RSS decreased when RAG1 and RAG2 were induced. In immunoblot analyses and gel supershift assays, we showed that KRC antisera react with the M_r 135,000 RSS-binding protein. We previously cloned KRC from a thymocyte expression library using ³²P-RSS as a ligand and showed that KRC fusion proteins bind specifically to the RSS and to the κB enhancer motif. The lymphoid expression and DNA-binding characteristics suggest that KRC may be involved in lymphocyte development.