Assembly of the Human R2TP Chaperone Complex

The Human R2TP complex is composed of hRvb1, hRvb2, RPAP3 and hPih1, and it is required for snoRNP biogenesis, PIKK signaling and RNA polymerase II assembly. Despite its important functions, much information on the structure of the human R2TP complex is still not available. In order to build up a foundation for the structural and functional studies of the human R2TP complex, I endeavored to reconstitute the complex in vitro. I successfully purified the human R2TP proteins, studied the interactions amongst these proteins, and attempted the complex reconstitution using different methods. Amongst the R2TP proteins, hRvb1 and hRvb2 could oligomerize together to form the hRvb1/hRvb2 complex, and RPAP3 and hPih1 could also interact to form the RPAP3/hPih1 complex. However, the interaction across the two sub-complexes of R2TP seemed to be rather weak, suggesting the successful formation of the complete R2TP complex might require help from extra assembly factors.M.Sc.2019-11-06 00:00:0