Document Detail


Nucleic acid polymerases use a general acid for nucleotidyl transfer.
MedLine Citation:
PMID:  19151724     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Nucleic acid polymerases catalyze the formation of DNA or RNA from nucleoside-triphosphate precursors. Amino acid residues in the active site of polymerases are thought to contribute only indirectly to catalysis by serving as ligands for the two divalent cations that are required for activity or substrate binding. Two proton-transfer reactions are necessary for polymerase-catalyzed nucleotidyl transfer: deprotonation of the 3'-hydroxyl nucleophile and protonation of the pyrophosphate leaving group. Using model enzymes representing all four classes of nucleic acid polymerases, we show that the proton donor to pyrophosphate is an active-site amino acid residue. The use of general acid catalysis by polymerases extends the mechanism of nucleotidyl transfer beyond that of the well-established two-metal-ion mechanism. The existence of an active-site residue that regulates polymerase catalysis may permit manipulation of viral polymerase replication speed and/or fidelity for virus attenuation and vaccine development.
Authors:
Christian Castro; Eric D Smidansky; Jamie J Arnold; Kenneth R Maksimchuk; Ibrahim Moustafa; Akira Uchida; Matthias Götte; William Konigsberg; Craig E Cameron
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2009-01-18
Journal Detail:
Title:  Nature structural & molecular biology     Volume:  16     ISSN:  1545-9985     ISO Abbreviation:  Nat. Struct. Mol. Biol.     Publication Date:  2009 Feb 
Date Detail:
Created Date:  2009-02-04     Completed Date:  2009-02-23     Revised Date:  2010-12-03    
Medline Journal Info:
Nlm Unique ID:  101186374     Medline TA:  Nat Struct Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  212-8     Citation Subset:  IM    
Affiliation:
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802, USA.
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / metabolism*
Catalysis
Manganese / metabolism
Models, Molecular
Nucleic Acids / metabolism
Nucleotidyltransferases / chemistry,  metabolism*
Protons
Viruses / enzymology*
Grant Support
ID/Acronym/Agency:
AI45818/AI/NIAID NIH HHS; R01 AI045818-10/AI/NIAID NIH HHS; R01 GM063276-08/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Nucleic Acids; 0/Protons; 7439-96-5/Manganese; EC 2.7.7.-/Nucleotidyltransferases
Comments/Corrections

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