Full-length dengue virus RNA-dependent RNA polymerase-RNA/DNA complexes

Stoichiometries, intrinsic affinities, cooperativities, base, and conformational specificities

Michal R. Szymanski, Maria J. Jezewska, Pawel Bujalowski, Cecile Bussetta, Mengyi Ye, Kyung Choi, Wlodzimierz Bujalowski

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Fundamental aspects of interactions of the Dengue virus type 3 full-length polymerase with the single-stranded and double-stranded RNA and DNA have been quantitatively addressed. The polymerase exists as a monomer with an elongated shape in solution. In the absence of magnesium, the total site size of the polymerase-ssRNA complex is 26 ± 2 nucleotides. In the presence of Mg 2+, the site size increases to 29 ± 2 nucleotides, indicating that magnesium affects the enzyme global conformation. The enzyme shows a preference for the homopyrimidine ssRNAs. Positive cooperativity in the binding to homopurine ssRNAs indicates that the type of nucleic acid base dramatically affects the enzyme orientation in the complex. Both the intrinsic affinity and the cooperative interactions are accompanied by a net ion release. The polymerase binds the dsDNA with an affinity comparable with the ssRNAs affinity, indicating that the binding site has an open conformation in solution. The lack of detectable dsRNA or dsRNA-DNA hybrid affinities indicates that the entry to the binding site is specific for the sugar-phosphate backbone and/or conformation of the duplex.

Original languageEnglish (US)
Pages (from-to)33095-33108
Number of pages14
JournalJournal of Biological Chemistry
Volume286
Issue number38
DOIs
StatePublished - Sep 23 2011

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RNA Replicase
Dengue Virus
Viruses
Stoichiometry
Conformations
RNA
Magnesium
DNA
Enzymes
Nucleotides
Binding Sites
Sugar Phosphates
Double-Stranded RNA
Nucleic Acids
Monomers
Ions

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Full-length dengue virus RNA-dependent RNA polymerase-RNA/DNA complexes : Stoichiometries, intrinsic affinities, cooperativities, base, and conformational specificities. / Szymanski, Michal R.; Jezewska, Maria J.; Bujalowski, Pawel; Bussetta, Cecile; Ye, Mengyi; Choi, Kyung; Bujalowski, Wlodzimierz.

In: Journal of Biological Chemistry, Vol. 286, No. 38, 23.09.2011, p. 33095-33108.

Research output: Contribution to journalArticle

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