Structural explanation for the role of Mn2+ in the activity of Φ6 RNA-dependent RNA polymerase

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  2. Dr Paula Salgado
Author(s)Poranen MM, Salgado PS, Koivunen MRL, Wright S, Bamford DH, Stuart DI, Grimes JM
Publication type Article
JournalNucleic Acids Research
Year2008
Volume36
Issue20
Pages6633-6644
ISSN (print)0305-1048
ISSN (electronic)1362-4962
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The biological role of manganese (Mn(2+)) has been a long-standing puzzle, since at low concentrations it activates several polymerases whilst at higher concentrations it inhibits. Viral RNA polymerases possess a common architecture, reminiscent of a closed right hand. The RNA-dependent RNA polymerase (RdRp) of bacteriophage 6 is one of the best understood examples of this important class of polymerases. We have probed the role of Mn(2+) by biochemical, biophysical and structural analyses of the wild-type enzyme and of a mutant form with an altered Mn(2+)-binding site (E491 to Q). The E491Q mutant has much reduced affinity for Mn(2+), reduced RNA binding and a compromised elongation rate. Loss of Mn(2+) binding structurally stabilizes the enzyme. These data and a re-examination of the structures of other viral RNA polymerases clarify the role of manganese in the activation of polymerization: Mn(2+) coordination of a catalytic aspartate is necessary to allow the active site to properly engage with the triphosphates of the incoming NTPs. The structural flexibility caused by Mn(2+) is also important for the enzyme dynamics, explaining the requirement for manganese throughout RNA polymerization.
PublisherOxford University Press
URLhttp://dx.doi.org/10.1093/nar/gkn632
DOI10.1093/nar/gkn632
PubMed id18940872
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