title: Preannealing of poly(A) template and oligo(dT) primer is not required for hepatitis C virus RNA-dependent RNA polymerase activity.
abstract: Preannealed homopolymeric DNAs or RNAs are often used as templates and/or primers to characterize activities of DNA or RNA-dependent RNA polymerases. Based on the calculated melting temperatures (T(m) values), however, poly(A)/oligo(dT(12-18)) is not expected to form stable duplexes. To determine this, we compared the enzymatic activity of hepatitis C virus polymerase using poly(A)/oligo(dT(12)) that were or were not preannealed. No significant differences were observed. These results suggest that it is not necessary to perform preannealing reactions for poly(A) and oligo(dT(12)), making it possible to characterize mechanism of inhibition of NS5B inhibitors against either template RNA poly(A) or primer oligo(dT(12)) independently.
authors.affiliation: Department of Virology, Bristol Myers Squibb Pharmaceutical Research Institute, Wallingford, CT 06067, USA. ying-kai.wang@bms.com
authors.forename: Ying-Kai
authors.initials: YK
authors.lastname: Wang
mesh_terms: D017931:DNA Primers / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D016174:Hepacivirus / Q000201:enzymology*; D011061:Poly A / Q000235:genetics / Q000378:metabolism*; D011071:Poly T / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013698:Templates, Genetic; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase
chemical_list: D017931:DNA Primers; D004791:Enzyme Inhibitors; D011061:Poly A; D011071:Poly T; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase