Identification of allosteric inhibitors blocking the hepatitis C virus polymerase NS5B in the RNA synthesis initiation step. - Archive ouverte HAL Access content directly
Journal Articles Antiviral Research Year : 2009

Identification of allosteric inhibitors blocking the hepatitis C virus polymerase NS5B in the RNA synthesis initiation step.

Stéphane Betzi
Cécilia Eydoux
Cécile Bussetta
  • Function : Author
Marilyne Blemont
  • Function : Author
Pieter Leyssen
  • Function : Author
Claire Debarnot
  • Function : Author
Mohamed Ben-Rahou
  • Function : Author
Jacques Haiech
  • Function : Author
Marcel Hibert
David S. Grierson
  • Function : Author
  • PersonId : 846963
Jean-Louis Romette
  • Function : Author
Jean-Claude Guillemot
  • Function : Author
Johan Neyts
Karine Alvarez
Xavier Morelli
Hélène Dutartre
  • Function : Author
Bruno Canard

Abstract

Hepatitis C virus (HCV) RNA-dependent RNA polymerase NS5B constitutes a target of choice for the development of anti-HCV drugs. Although many small molecules have been identified as allosteric inhibitors of NS5B, very few are active in clinical applications. We have screened 17,000 compounds in an enzymatic assay involving the purified NS5B in order to increase the therapeutic arsenal. We hoped to shed some light on the precise mechanism of RNA synthesis. We succeeded in isolating a series of 21 original inhibitors of the RNA synthesis by NS5B. Four of these non-nucleoside inhibitors (NNIs) could be mapped to the known binding site called 'B' as judged by the decrease in their inhibition potency when assayed with a 'B' site mutant, M423T NS5B. Incidentally, our in silico model pointed to Y477 as a key residue for inhibitor binding. In vitro, Y477F mutant loses its sensitivity to the newly discovered inhibitors but is unable to extend primers during the elongation phase. Our results demonstrate that elements of the 'B' site are involved in the conformational changes required in the switch between the different RNA synthesis steps and that compounds targeting this site could lock the enzyme in its initiation phase.

Dates and versions

hal-00418783 , version 1 (21-09-2009)

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Cite

Stéphane Betzi, Cécilia Eydoux, Cécile Bussetta, Marilyne Blemont, Pieter Leyssen, et al.. Identification of allosteric inhibitors blocking the hepatitis C virus polymerase NS5B in the RNA synthesis initiation step.. Antiviral Research, 2009, 84 (1), pp.48-59. ⟨10.1016/j.antiviral.2009.06.009⟩. ⟨hal-00418783⟩
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