title: Primuline derivatives that mimic RNA to stimulate hepatitis C virus NS3 helicase-catalyzed ATP hydrolysis.
abstract: ATP hydrolysis fuels the ability of helicases and related proteins to translocate on nucleic acids and separate base pairs. As a consequence, nucleic acid binding stimulates the rate at which a helicase catalyzes ATP hydrolysis. In this study, we searched a library of small molecule helicase inhibitors for compounds that stimulate ATP hydrolysis catalyzed by the hepatitis C virus (HCV) NS3 helicase, which is an important antiviral drug target. Two compounds were found that stimulate HCV helicase-catalyzed ATP hydrolysis, both of which are amide derivatives synthesized from the main component of the yellow dye primuline. Both compounds possess a terminal pyridine moiety, which was critical for stimulation. Analogs lacking a terminal pyridine inhibited HCV helicase catalyzed ATP hydrolysis. Unlike other HCV helicase inhibitors, the stimulatory compounds differentiate between helicases isolated from various HCV genotypes and related viruses. The compounds only stimulated ATP hydrolysis catalyzed by NS3 purified from HCV genotype 1b. They inhibited helicases from other HCV genotypes (e.g. 1a and 2a) or related flaviviruses (e.g. Dengue virus). The stimulatory compounds interacted with HCV helicase in the absence of ATP with dissociation constants of about 2 μM. Molecular modeling and site-directed mutagenesis studies suggest that the stimulatory compounds bind in the HCV helicase RNA-binding cleft near key residues Arg-393, Glu-493, and Ser-231.
authors.affiliation: Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin 53211, USA.
authors.forename: Noreena L
authors.initials: NL
authors.lastname: Sweeney
authors.forename: William R
authors.initials: WR
authors.lastname: Shadrick
authors.forename: Sourav
authors.initials: S
authors.lastname: Mukherjee
authors.forename: Kelin
authors.initials: K
authors.lastname: Li
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Frankowski
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Schoenen
authors.forename: David N
authors.initials: DN
authors.lastname: Frick
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry* / Q000378:metabolism; D016174:Hepacivirus / Q000201:enzymology* / Q000235:genetics; D006868:Hydrolysis; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D020365:RNA Helicases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral; D013844:Thiazoles / Q000737:chemistry*; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D012367:RNA, Viral; D013844:Thiazoles; D014764:Viral Proteins; C035646:primuline; D000255:Adenosine Triphosphate; D020365:RNA Helicases
keywords: ATPases; Chemical Biology; Drug Discovery; High Throughput Screening (HTS); Molecular Motors; RNA Helicase