Selection of the same mutation in the U69 protein kinase gene of human herpesvirus-6 after prolonged exposure to ganciclovir in vitro and in vivo.
After serial passage in the presence of increasing concentrations of ganciclovir (GCV) in vitro, a human herpesvirus-6 (HHV-6) mutant exhibiting a decreased sensitivity to the drug was isolated. Analysis of drug susceptibility showed that the IC(50) of this mutant was 24-, 52- and 3-fold higher than that of the wild-type (wt) IC(50) in the case of GCV, cidofovir and foscarnet, respectively. Genotypic analysis showed two single nucleotide changes as compared to the wild-type: an A-->G substitution of the U69 protein kinase (PK) gene resulted in an M(318)V amino acid substitution and the other change, located in the C-terminal part of the U38 gene, resulted in an A(961)V amino acid substitution within the DNA polymerase. The M(318)V change was located within the consensus sequence DISPMN of the putative catalytic domain VI of the PK. This change was homologous to the M(460)V and M(460)I changes that had been reported previously within the consensus sequence DITPMN of the human cytomegalovirus (HCMV) UL97 PK and associated with the resistance of HCMV to GCV. The M(318)V change was also detected by PCR in HHV-6-infected PBMCs from an AIDS patient who had been treated with GCV for a long period of time and exhibited a clinically GCV-resistant HCMV infection. These findings provide strong circumstantial evidence that the M(318)V change of the PK gene is associated with resistance to GCV and raise the question of cross resistance to this drug among different betaherpesviruses.
Manichanh
Chaysavanh
C
Laboratoire de Virologie, UPRES EA 2387, CERVI1, Laboratoire de Génétique Moléculaire, Service de Biochimie Médicale2 and Service des Maladies Infectieuses et Tropicales3, Groupe Hospitalier Pitié-Salpêtrière, 83 Bld de l'Hôpital, 75651 Paris Cedex 13, France.
Olivier-Aubron
Camille
C
Laboratoire de Virologie, UPRES EA 2387, CERVI1, Laboratoire de Génétique Moléculaire, Service de Biochimie Médicale2 and Service des Maladies Infectieuses et Tropicales3, Groupe Hospitalier Pitié-Salpêtrière, 83 Bld de l'Hôpital, 75651 Paris Cedex 13, France.
Lagarde
Jean-Pierre
JP
Laboratoire de Virologie, UPRES EA 2387, CERVI1, Laboratoire de Génétique Moléculaire, Service de Biochimie Médicale2 and Service des Maladies Infectieuses et Tropicales3, Groupe Hospitalier Pitié-Salpêtrière, 83 Bld de l'Hôpital, 75651 Paris Cedex 13, France.
Aubin
Jean-Thierry
JT
Laboratoire de Virologie, UPRES EA 2387, CERVI1, Laboratoire de Génétique Moléculaire, Service de Biochimie Médicale2 and Service des Maladies Infectieuses et Tropicales3, Groupe Hospitalier Pitié-Salpêtrière, 83 Bld de l'Hôpital, 75651 Paris Cedex 13, France.
Bossi
Phillipe
P
Laboratoire de Virologie, UPRES EA 2387, CERVI1, Laboratoire de Génétique Moléculaire, Service de Biochimie Médicale2 and Service des Maladies Infectieuses et Tropicales3, Groupe Hospitalier Pitié-Salpêtrière, 83 Bld de l'Hôpital, 75651 Paris Cedex 13, France.
Gautheret-Dejean
Agnès
A
Laboratoire de Virologie, UPRES EA 2387, CERVI1, Laboratoire de Génétique Moléculaire, Service de Biochimie Médicale2 and Service des Maladies Infectieuses et Tropicales3, Groupe Hospitalier Pitié-Salpêtrière, 83 Bld de l'Hôpital, 75651 Paris Cedex 13, France.
Huraux
Jean-Marie
JM
Laboratoire de Virologie, UPRES EA 2387, CERVI1, Laboratoire de Génétique Moléculaire, Service de Biochimie Médicale2 and Service des Maladies Infectieuses et Tropicales3, Groupe Hospitalier Pitié-Salpêtrière, 83 Bld de l'Hôpital, 75651 Paris Cedex 13, France.
Agut
Henri
H
Laboratoire de Virologie, UPRES EA 2387, CERVI1, Laboratoire de Génétique Moléculaire, Service de Biochimie Médicale2 and Service des Maladies Infectieuses et Tropicales3, Groupe Hospitalier Pitié-Salpêtrière, 83 Bld de l'Hôpital, 75651 Paris Cedex 13, France.
D017088:AIDS-Related Opportunistic Infections / Q000821:virology; D000328:Adult; D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000494:pharmacology* / Q000627:therapeutic use; D002460:Cell Line; D003587:Cytomegalovirus / Q000187:drug effects; D003586:Cytomegalovirus Infections / Q000150:complications / Q000188:drug therapy / Q000821:virology; D024882:Drug Resistance, Viral / Q000235:genetics*; D005077:Exanthema Subitum / Q000188:drug therapy / Q000821:virology*; D005260:Female; D015774:Ganciclovir / Q000494:pharmacology* / Q000627:therapeutic use; D015654:Herpesvirus 6, Human / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests / Q000379:methods; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D011494:Protein Kinases / Q000235:genetics*; D017422:Sequence Analysis, DNA
D000998:Antiviral Agents; D011494:Protein Kinases; D015774:Ganciclovir