Noncatalytic Ions Direct the RNA-Dependent RNA Polymerase of Bacterial Double-Stranded RNA Virus phi 6 from De Novo Initiation to Elongation

作者：Wright Sam; Poranen Minna M; Bamford Dennis H; Stuart David I; Grimes Jonathan M^*

来源：Journal of Virology, 2012, 86(5): 2837-2849.

DOI：10.1128/JVI.05168-11

RNA-dependent RNA polymerases (RdRps) are key to the replication of RNA viruses. A common divalent cation binding site, distinct from the positions of catalytic ions, has been identified in many viral RdRps. We have applied biochemical, biophysical, and structural approaches to show how the RdRp from bacteriophage phi 6 uses the bound noncatalytic Mn2+ to facilitate the displacement of the C-terminal domain during the transition from initiation to elongation. We find that this displacement releases the noncatalytic Mn2+, which must be replaced for elongation to occur. By inserting a dysfunctional Mg2+ at this site, we captured two nucleoside triphosphates within the active site in the absence of Watson-Crick base pairing with template and mapped movements of divalent cations during preinitiation. These structures refine the pathway from preinitiation through initiation to elongation for the RNA-dependent RNA polymerization reaction, explain the role of the noncatalytic divalent cation in phi 6 RdRp, and pinpoint the previously unresolved Mn2+-dependent step in replication.

• 出版日期2012-3