title: Mutations in the non-nucleoside binding-pocket interfere with the multi-nucleoside resistance phenotype.
abstract: OBJECTIVE
  To investigate the genotypic and phenotypic effects of in vitro resistance selection with lamivudine and/or the second generation non-nucleoside reverse transcriptase inhibitor (NNRTI) quinoxaline HBY097 using HIV-1 isolates carrying the multi-nucleoside resistance pattern linked to the Q151M mutation.
  
  
  METHODS
  Virus strains were selected in C8166 cells in the presence of increasing concentrations of lamivudine or HBY097. In parallel control experiments, the virus was cultured in C8166 cells in the absence of drugs. The entire reverse transcriptase encoding region was amplified using polymerase chain reaction and was subsequently sequenced. Antiviral activities of drugs were evaluated in C8166 cells.
  
  
  RESULTS
  High-level resistant viruses were selected rapidly in the presence of lamivudine and quinoxaline (less than 10 passages). The multi-nucleoside resistance mutations were stable during in vitro resistance selection. Lamivudine elicited the acquisition of the M184I mutation. Phenotypic resistance to all nucleoside-analog reverse transcriptase inhibitors (NRTIs) was increased when M184I was added to the multi-nucleoside resistance background in the absence of NNRTI-resistance mutations. In most cases of HBY097 resistance selection, at least two mutations associated with NNRTI resistance resulted in high-level NNRTI resistance. The NNRTI resistance-related mutations partially reversed the phenotypic resistance to most NRTIs, except to abacavir. The addition of the M184I mutation to the NNRTI-multi-nucleoside resistance set abolished this antagonizing effect for didanosine, zalcitabine and lamivudine, but further potentiated the phenotypic reversal for zidovudine and stavudine.
  
  
  CONCLUSIONS
  Changes in the non-nucleoside binding pocket must affect the conformation of residues at the dNTP binding site, and can result in a partial phenotypic reversal of the multi-nucleoside resistance phenotype.
authors.affiliation: Rega Institute for Medical Research and University Hospitals, Katholieke Universiteit Leuven, Belgium.
authors.forename: K
authors.initials: K
authors.lastname: Van Laethem
authors.forename: M
authors.initials: M
authors.lastname: Witvrouw
authors.forename: C
authors.initials: C
authors.lastname: Pannecouque
authors.forename: B
authors.initials: B
authors.lastname: Van Remoortel
authors.forename: J C
authors.initials: JC
authors.lastname: Schmit
authors.forename: R
authors.initials: R
authors.lastname: Esnouf
authors.forename: J P
authors.initials: JP
authors.lastname: Kleim
authors.forename: J
authors.initials: J
authors.lastname: Balzarini
authors.forename: J
authors.initials: J
authors.lastname: Desmyter
authors.forename: E
authors.initials: E
authors.lastname: De Clercq
authors.forename: A M
authors.initials: AM
authors.lastname: Vandamme
mesh_terms: D019380:Anti-HIV Agents / Q000378:metabolism / Q000494:pharmacology*; D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology*; D001665:Binding Sites; D004352:Drug Resistance, Microbial; D018432:Drug Resistance, Multiple / Q000235:genetics*; D005838:Genotype; D015658:HIV Infections / Q000188:drug therapy / Q000821:virology*; D054303:HIV Reverse Transcriptase / Q000235:genetics*; D015497:HIV-1 / Q000145:classification / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D019259:Lamivudine / Q000378:metabolism / Q000494:pharmacology*; D009154:Mutation; D009705:Nucleosides / Q000378:metabolism / Q000494:pharmacology; D010641:Phenotype; D011810:Quinoxalines / Q000378:metabolism / Q000494:pharmacology*; D018894:Reverse Transcriptase Inhibitors / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D019380:Anti-HIV Agents; D000998:Antiviral Agents; D009705:Nucleosides; D011810:Quinoxalines; D018894:Reverse Transcriptase Inhibitors; D019259:Lamivudine; D054303:HIV Reverse Transcriptase; C093110:HBY 097