The two key factors for the generation of the diverse antigen receptor repertoire in the human adaptive immune system are the products of the recombination activating genes 1 and 2 (RAG1 and RAG2). The identification of their homologs (spRAG1L and spRAG2L, respectively) in the genome of the purple sea urchin (Strongylocentrotus purpuratus) was unexpected as there is no evidence thus far of an adaptive immune system in invertebrates (including echinoderms), i.e. they lack at least one of its hallmarks, a diversified antigen receptor repertoire. During this fiscal year we continued our studies to determine the DNA substrate and the enzymatic activity of the SpRag1L/SpRag2L complex using hybrid proteins in which the DNA-binding domains of Rag1 and SpRag1L have been swapped. We determined that our chimeric proteins maintained their ability to interact with the respective Rag2 partner protein, and initiated experiments to test their function in transient recombination assays in vivo. Furthermore, our hybrid proteins were also purified as recombinant proteins from E coli, and their biochemical characterization is ongoing. Our studies have major implications for the current model of how adaptive immunity evolved in jawed vertebrates, and will help to illuminate conserved features of how V(D)J recombination is tightly controlled to avoid potentially dangerous modifications of the genome.
| Fugmann, Sebastian D (2011) RAG-2 unleashed: lymphocytes beware. Immunity 34:137-9 |
| Litman, Gary W; Rast, Jonathan P; Fugmann, Sebastian D (2010) The origins of vertebrate adaptive immunity. Nat Rev Immunol 10:543-53 |
| Fugmann, Sebastian D (2010) The origins of the Rag genes--from transposition to V(D)J recombination. Semin Immunol 22:10-6 |
