Dynamics on multiple timescales in the RNA-directed RNA polymerase from the cystovirus phi6.
The de novo initiating RNA-directed RNA polymerase (RdRP), P2, forms the central machinery in the infection cycle of the bacteriophage phi6 by performing the dual tasks of replication and transcription of the double-stranded RNA genome in the host cell. By measurement and quantitative analysis of multiple-quantum spin-relaxation data for the delta1 positions of Ile residues that are distributed over the 3D-fold of P2, we find that the enzyme is dynamic both on the fast (ps-ns) and slow (micros-ms) timescales. The characteristics of several motional modes including those that coincide with the catalytic timescale (500-800/s) are altered in the presence of substrate analogs and single-stranded RNA templates. These studies reveal the plasticity of this finely tuned molecular machine and represent a first step towards linking structural information available from a host of crystal structures to catalytic mechanisms and timescales obtained from the measurements of kinetics for homologous systems in solution.
Ren
Zhen
Z
Department of Chemistry, The City College of New York, New York, NY 10031, USA.
Wang
Hsin
H
Ghose
Ranajeet
R
D017126:Bacteriophage phi 6 / Q000201:enzymology*; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D007532:Isoleucine / Q000737:chemistry; D056004:Molecular Dynamics Simulation; D019906:Nuclear Magnetic Resonance, Biomolecular; D012313:RNA / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry* / Q000378:metabolism
D007532:Isoleucine; C004805:5'-guanylylmethylenebisphosphonate; D012313:RNA; D006160:Guanosine Triphosphate; D012324:RNA-Dependent RNA Polymerase
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