Single amino acid changes in the DNA polymerase confer foscarnet resistance and slow-growth phenotype, while mutations in the UL97-encoded phosphotransferase confer ganciclovir resistance in three double-resistant human cytomegalovirus strains recovered from patients with AIDS
| ISSN: | 1098-5514 0022-538X |
|---|---|
| DOI: | 10.1128/jvi.70.3.1390-1395.1996 |
| Přístupová URL adresa: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d5c0fe328c13771d01d49a7f5b990340 https://doi.org/10.1128/jvi.70.3.1390-1395.1996 |
| Rights: | OPEN |
| Přírůstkové číslo: | edsair.doi.dedup.....d5c0fe328c13771d01d49a7f5b990340 |
| Autor: | Antonella Sarasini, Fausto Baldanti, Mark R. Underwood, Sylvia C. Stanat, Enrico Silini, Sunwen Chou, Karen K. Biron, Giuseppe Gerna |
| Rok vydání: | 1996 |
| Předmět: |
Foscarnet
Human cytomegalovirus Ganciclovir DNA polymerase viruses Molecular Sequence Data Immunology Population Cytomegalovirus DNA-Directed DNA Polymerase medicine.disease_cause Microbiology law.invention law Virology medicine Humans Amino Acid Sequence Phosphorylation education Gene Mutation education.field_of_study AIDS-Related Opportunistic Infections Base Sequence biology Drug Resistance Microbial medicine.disease Molecular biology Drug Resistance Multiple Phosphotransferases (Alcohol Group Acceptor) Phenotype Insect Science Cytomegalovirus Infections biology.protein Recombinant DNA Research Article medicine.drug |
| Zdroj: | Scopus-Elsevier |
| ISSN: | 1098-5514 0022-538X |
| DOI: | 10.1128/jvi.70.3.1390-1395.1996 |
| Popis: | Three human cytomegalovirus (HCMV) strains (VR4760, VR4955, and VR5120) showing double resistance to ganciclovir (GCV) and foscarnet (PFA) were isolated from three patients with AIDS who underwent multiple sequential courses of therapy with GCV and PFA (A. Sarasini, F. Baldanti, M. Furione, E. Percivalle, R. Brerra, M. Barbi, and G. Gerna, J. Med. Virol., 47:237-244, 1995). We previously demonstrated that the three strains were genetically unrelated and that each of them was present as a single viral population in vivo. Thus, in each of the three cases, a single viral strain was resistant to both GCV and PFA. In the present paper, we report the characterization of the molecular bases of the double resistance and demonstrate that the PFA resistance is associated with a slower replication of HCMV strains in cell cultures. Sequencing of the UL97 and UL54 genes, GCV anabolism assays, and marker transfer experiments showed that GCV resistance was due to single amino acid changes in the UL97 gene product (VR4760, Met-460 --> Ile; VR4955, Ala-594 --> Val; VR5120, Leu595 --> Ser), while single amino acid changes in domain II of the DNA polymerase (VR4760 and VR5120, Val-715 --> Met; VR4955, Thr-700 --> Ala) were responsible for both the PFA resistance and the slow-growth phenotype. Thus, in these three cases, double resistance to GCV and PFA was not due to a single mutation conferring cross-resistance or to the presence of a mixture of strains with different drug susceptibilities. The HCMV DNA polymerase recombinant strains carrying the mutations conferring PFA resistance were sensitive to GCV and (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine (HPMPC). In addition, the same UL54 mutations were responsible for the slow growth of the clinical isolates, since the recombinant strains showed a marked delay in immediate-early antigen plaque formation and a reduction of infectious virus yield compared with AD169, from which they were derived. These results may have some important implications for the successful isolation, propagation, and characterization of PFA-resistant strains from clinical samples containing mixed viral populations. |
| Databáze: | OpenAIRE |
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