title: The polymerase L528M mutation cooperates with nucleotide binding-site mutations, increasing hepatitis B virus replication and drug resistance.
abstract: After receiving lamivudine for 3 years to treat chronic hepatitis B, 67-75% of patients develop B-domain L528M, C-domain M552I, or M552V mutations in the HBV polymerase that render hepatitis B virus (HBV) drug-resistant. The aim of this study was to evaluate the influence of these mutations on viral replication and resistance to antiviral agents. We investigated the replication fitness and susceptibility of the wild-type and five mutant HBVs (L528M, M552I, M552V, L528M/M552I, and L528M/M552V) to 11 compounds [lamivudine, adefovir, entecavir (BMS-200475) (+)-BCH-189 (+/-)-FTC (racivir) (-)-FTC (emtricitabine) (+)-FTC, L-D4FC, L-FMAU (clevudine), D-DAPD, and (-)-carbovir] by transfecting HBV DNA into hepatoma cells and monitoring viral products by Southern blotting. The replication competency of the single C-domain mutants M552I and M552V was markedly decreased compared with that of wild-type HBV. However, addition of the B-domain mutation L528M restored replication competence. Only adefovir and entecavir were effective against all five HBV mutants, and higher doses of these compounds were necessary to inhibit the double mutants compared with the single mutants. The B-domain mutation (L528M) of HBV polymerase not only restores the replication competence of C-domain mutants, but also increases resistance to nucleoside analogues.
authors.affiliation: Department of Gastroenterology, Faculty of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan.
authors.forename: S K
authors.initials: SK
authors.lastname: Ono
authors.forename: N
authors.initials: N
authors.lastname: Kato
authors.forename: Y
authors.initials: Y
authors.lastname: Shiratori
authors.forename: J
authors.initials: J
authors.lastname: Kato
authors.forename: T
authors.initials: T
authors.lastname: Goto
authors.forename: R F
authors.initials: RF
authors.lastname: Schinazi
authors.forename: F J
authors.initials: FJ
authors.lastname: Carrilho
authors.forename: M
authors.initials: M
authors.lastname: Omata
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D001665:Binding Sites; D002478:Cells, Cultured; D004351:Drug Resistance; D006515:Hepatitis B virus / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D006801:Humans; D009154:Mutation; D009711:Nucleotides / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000235:genetics*; D014779:Virus Replication
chemical_list: D000998:Antiviral Agents; D009711:Nucleotides; D012194:RNA-Directed DNA Polymerase