title: Contrasting effect of new HCMV pUL54 mutations on antiviral drug susceptibility: Benefits and limits of 3D analysis.
abstract: Human cytomegalovirus (HCMV) resistance to antiviral drugs is a major drawback of repeated or long-duration treatment in immunocompromised patients. Resistance testing is usually performed by genotypic assays. For accurate interpretation of these assays, the role of new mutations in HCMV resistance has to be assessed. Two previously unknown UL54 single point mutations (D515Y and V787A) were characterized for phenotypic drug-resistance by marker transfer analysis using bacterial artificial chromosome (BAC) mutagenesis. Increases in 50% inhibitory concentrations of ganciclovir and foscarnet were found for both mutated recombinant strains showing that mutations D515Y and V787A induce resistance to both antivirals. Importantly, none of those impacted the viral growth kinetics. For a better understanding of their molecular resistance mechanisms, a 3D homology model was used to localize the mutated amino-acids in functional domains of UL54 and predict their impact on UL54 function and resistance. However, 3D homology model analysis has limits and phenotypic characterization using BAC-HCMV is still essential to measure the role of unknown mutations.
authors.affiliation: Univ. Limoges, UMR, 1092, Limoges, France; INSERM, UMR, 1092, Limoges, France; CHU Limoges, Laboratoire de Bactériologie-Virologie-Hygiène, National Reference Center for Cytomegaloviruses (NRC), Limoges, France.
authors.forename: D
authors.initials: D
authors.lastname: Andouard
authors.affiliation: CHU Saint Louis, Laboratoire de Bactériologie-Virologie, NRC-Associated Laboratory, Paris, France.
authors.forename: M-C
authors.initials: MC
authors.lastname: Mazeron
authors.affiliation: Univ. Limoges, UMR, 1092, Limoges, France; INSERM, UMR, 1092, Limoges, France; CHU Limoges, Laboratoire de Bactériologie-Virologie-Hygiène, National Reference Center for Cytomegaloviruses (NRC), Limoges, France.
authors.forename: G
authors.initials: G
authors.lastname: Ligat
authors.affiliation: Univ. Limoges, UMR, 1092, Limoges, France; INSERM, UMR, 1092, Limoges, France; CHU Limoges, Laboratoire de Bactériologie-Virologie-Hygiène, National Reference Center for Cytomegaloviruses (NRC), Limoges, France.
authors.forename: A
authors.initials: A
authors.lastname: Couvreux
authors.affiliation: CHU Lyon, Service de Transplantation rénale, Hôpital Edouard Herriot, Lyon, France.
authors.forename: C
authors.initials: C
authors.lastname: Pouteil-Noble
authors.affiliation: CHU Lyon, Service de Transplantation rénale, Hôpital Edouard Herriot, Lyon, France.
authors.forename: R
authors.initials: R
authors.lastname: Cahen
authors.affiliation: CHU Bichat, Service de Maladies infectieuses et Tropicales, Paris, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Yasdanpanah
authors.affiliation: Univ. Limoges, UMR, 1092, Limoges, France; INSERM, UMR, 1092, Limoges, France; CHU Limoges, Laboratoire de Bactériologie-Virologie-Hygiène, National Reference Center for Cytomegaloviruses (NRC), Limoges, France.
authors.forename: M
authors.initials: M
authors.lastname: Deering
authors.affiliation: Department of Infectious Diseases, Lille School of Medicine, Tourcoing Hospital, Tourcoing, France.
authors.forename: N
authors.initials: N
authors.lastname: Viget
authors.affiliation: Univ. Limoges, UMR, 1092, Limoges, France; INSERM, UMR, 1092, Limoges, France; CHU Limoges, Laboratoire de Bactériologie-Virologie-Hygiène, National Reference Center for Cytomegaloviruses (NRC), Limoges, France.
authors.forename: S
authors.initials: S
authors.lastname: Alain
authors.affiliation: Univ. Limoges, UMR, 1092, Limoges, France; INSERM, UMR, 1092, Limoges, France; CHU Limoges, Laboratoire de Bactériologie-Virologie-Hygiène, National Reference Center for Cytomegaloviruses (NRC), Limoges, France. Electronic address: sebastien.hantz@unilim.fr.
authors.forename: S
authors.initials: S
authors.lastname: Hantz
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D022202:Chromosomes, Artificial, Bacterial; D003587:Cytomegalovirus / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D003596:Cytosine / Q000494:pharmacology; D004279:DNA, Viral / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000235:genetics*; D024882:Drug Resistance, Viral; D017245:Foscarnet / Q000494:pharmacology; D015774:Ganciclovir / Q000494:pharmacology; D006801:Humans; D008958:Models, Molecular; D016296:Mutagenesis; D063065:Organophosphonates / Q000494:pharmacology; D010641:Phenotype; D017354:Point Mutation; D000072417:Protein Domains; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics*
chemical_list: D000998:Antiviral Agents; D004279:DNA, Viral; D063065:Organophosphonates; C109367:UL54 protein, Human herpesvirus 5; D014764:Viral Proteins; D017245:Foscarnet; D003596:Cytosine; D004259:DNA-Directed DNA Polymerase; D015774:Ganciclovir
keywords: 3D-model; BAC; Cytomegalovirus; Foscarnet; Ganciclovir; Resistance