Yuhang Zhang 1 , 8 , Tat Cheung Cheng 2 , 8 , Guangrui Huang 3 , Qingyi Lu 3 , Marius D. Surleac 4 , Jeffrey D. Mandell 1 , Pierre Pontarotti 5 , 6 , Andrei J. Petrescu 4 , Anlong Xu 3 , 7 , * , Yong Xiong 2 , * , David G. Schatz 1 , *
10 April 2019
Domestication of a transposon to give rise to the RAG1/RAG2 recombinase and V(D)J recombination was a pivotal event in the evolution of the jawed vertebrate adaptive immune system. The evolutionary adaptations that transformed the ancestral RAG transposase into a RAG recombinase with appropriately regulated DNA cleavage and transposition activities are not understood. Here, beginning with cryo-electron microscopy structures of RAG’s evolutionary relative, the ProtoRAG transposase from amphioxus, we identify amino acid residues and domains whose acquisition or loss underpins RAG’s propensity for coupled cleavage, preference for asymmetric DNA substrates, and inability to perform transposition in cells. In particular, we identify two jawed-vertebrate-specific adaptations—arginine 848 in RAG1 and an acidic region in RAG2—that together suppress RAG-mediated transposition more than 1000-fold. Our findings reveal a two-tiered mechanism for suppression of RAG-mediated transposition, illuminate the forces at work during the evolution of V(D)J recombination, and provide insight into the principles governing transposon molecular domestication.