Resistance of Human Cytomegalovirus to the Benzimidazole L-Ribonucleoside Maribavir Maps to UL27

Gloria Komazin-Meredith, Roger G. Ptak, Brian T. Emmer, Leroy B. Townsend, John C. Drach

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

1-(β-D-Ribofuranosyl)-2,5,6-trichlorobenzimidazole (TCRB) and its 2-bromo analog, BDCRB, are potent and selective inhibitors of human cytomegalovirus (HCMV) DNA processing and packaging. Since they are readily metabolized in vivo, analogs were synthesized to improve biostability. One of these, 1-(β-L-ribofuranosyl)-2-isopropylamino-5,6-dichlorobenzimidazole (1263W94; maribavir), inhibits viral DNA synthesis and nuclear egress. Resistance to maribavir was mapped to UL97, and this viral kinase was shown to be a direct target of maribavir. In the present study, an HCMV strain resistant to TCRB and BDCRB was passaged in increasing concentrations of maribavir, and resistant virus was isolated. This strain (G2) grew at the same rate as the wild-type virus and was resistant to both BDCRB and maribavir. Resistance to BDCRB was expected, because the parent strain from which G2 was isolated was resistant due to known mutations in UL56 and UL89. However, no mutations were found in UL97 or other relevant open reading frames that could explain resistance to maribavir. Because sequencing of selected HCMV genes did not identify the resistance mutation, a cosmid library was made from G2, and a series of recombinant G2 wild-type viruses were constructed. Testing the recombinants for sensitivity to maribavir narrowed the locus of resistance to genes UL26 to UL32. Sequencing identified a single coding mutation in ORF UL27 (Leu335Pro) as the one responsible for resistance to maribavir. These results establish that UL27 is either directly or indirectly involved in the mechanism of action of maribavir. They also suggest that UL27 could play a role in HCMV DNA synthesis or egress of HCMV particles from the nucleus.

Original languageEnglish (US)
Pages (from-to)11499-11506
Number of pages8
JournalJournal of virology
Volume77
Issue number21
DOIs
StatePublished - Nov 1 2003

Fingerprint

ribonucleosides
Human herpesvirus 5
Ribonucleosides
benzimidazole
Cytomegalovirus
mutation
viruses
open reading frames
DNA
Mutation
synthesis
Viruses
packaging
Open Reading Frames
mechanism of action
phosphotransferases (kinases)
genes
maribavir
DNA Packaging
Cosmids

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Komazin-Meredith, Gloria ; Ptak, Roger G. ; Emmer, Brian T. ; Townsend, Leroy B. ; Drach, John C. / Resistance of Human Cytomegalovirus to the Benzimidazole L-Ribonucleoside Maribavir Maps to UL27. In: Journal of virology. 2003 ; Vol. 77, No. 21. pp. 11499-11506.
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Resistance of Human Cytomegalovirus to the Benzimidazole L-Ribonucleoside Maribavir Maps to UL27. / Komazin-Meredith, Gloria; Ptak, Roger G.; Emmer, Brian T.; Townsend, Leroy B.; Drach, John C.

In: Journal of virology, Vol. 77, No. 21, 01.11.2003, p. 11499-11506.

Research output: Contribution to journalArticle

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AU - Komazin-Meredith, Gloria

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AB - 1-(β-D-Ribofuranosyl)-2,5,6-trichlorobenzimidazole (TCRB) and its 2-bromo analog, BDCRB, are potent and selective inhibitors of human cytomegalovirus (HCMV) DNA processing and packaging. Since they are readily metabolized in vivo, analogs were synthesized to improve biostability. One of these, 1-(β-L-ribofuranosyl)-2-isopropylamino-5,6-dichlorobenzimidazole (1263W94; maribavir), inhibits viral DNA synthesis and nuclear egress. Resistance to maribavir was mapped to UL97, and this viral kinase was shown to be a direct target of maribavir. In the present study, an HCMV strain resistant to TCRB and BDCRB was passaged in increasing concentrations of maribavir, and resistant virus was isolated. This strain (G2) grew at the same rate as the wild-type virus and was resistant to both BDCRB and maribavir. Resistance to BDCRB was expected, because the parent strain from which G2 was isolated was resistant due to known mutations in UL56 and UL89. However, no mutations were found in UL97 or other relevant open reading frames that could explain resistance to maribavir. Because sequencing of selected HCMV genes did not identify the resistance mutation, a cosmid library was made from G2, and a series of recombinant G2 wild-type viruses were constructed. Testing the recombinants for sensitivity to maribavir narrowed the locus of resistance to genes UL26 to UL32. Sequencing identified a single coding mutation in ORF UL27 (Leu335Pro) as the one responsible for resistance to maribavir. These results establish that UL27 is either directly or indirectly involved in the mechanism of action of maribavir. They also suggest that UL27 could play a role in HCMV DNA synthesis or egress of HCMV particles from the nucleus.

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