Differential divalent cation requirements uncouple the assembly and catalytic reactions of human immunodeficiency virus type 1 integrase.

摘要: Previous in vitro analyses have shown that the human immunodeficiency virus type 1 (HIV-1) integrase uses either manganese or magnesium to assemble as a stable complex on donor substrate and catalyze strand transfer. We now demonstrate subsequent assembly, catalysis of both 3' end processing transfer requires divalent cation cofactor requirements for assembly can be functionally distinguished based ability utilize calcium cobalt, respectively. The different manifest by these processes are exploited uncouple catalysis, thus staging reaction. Staged assays then used conjunction with exonuclease III protection analysis investigate effects inhibitors each step Analysis series related demonstrates types compounds affect not catalytic process, therefore reconciling apparent disparate results obtained such using isolated preintegration complexes. These studies provide evidence distinct role implications identification characterization inhibitors.