Characterization of dengue virus resistance to brequinar in cell culture

Min Qing, Gang Zou, Qing Yin Wang, Hao Ying Xu, Hongping Dong, Zhiming Yuan, Pei-Yong Shi

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Brequinar is an inhibitor of dihydroorotate dehydrogenase, an enzyme that is required for de novo pyrimidine biosynthesis. Here we report that brequinar has activity against a broad spectrum of viruses. The compound not only inhibits flaviviruses (dengue virus, West Nile virus, yellow fever virus, and Powassan virus) but also suppresses a plus-strand RNA alphavirus (Western equine encephalitis virus) and a negative-strand RNA rhabdovirus (vesicular stomatitis virus). Using dengue virus serotype 2 (DENV-2) as a model, we found that brequinar suppressed the viral infection cycle mainly at the step of RNA synthesis. Supplementing the culture medium with pyrimidines (cytidine or uridine) but not purines (adenine or guanine) could be used to reverse the inhibitory effect of the compound. Continuous culturing of DENV-2 in the presence of brequinar generated viruses that were partially resistant to the inhibitor. Sequencing of the resistant viruses revealed two amino acid mutations: one mutation (M260V) located at a helix in the domain II of the viral envelope protein and another mutation (E802Q) located at the priming loop of the nonstructural protein 5 (NS5) polymerase domain. Functional analysis of the mutations suggests that the NS5 mutation exerts resistance through enhancement of polymerase activity. The envelope protein mutation reduced the efficiency of virion assembly/release; however, the mutant virus became less sensitive to brequinar inhibition at the step of virion assembly/release. Taken together, the results indicate that (i) brequinar blocks DENV RNA synthesis through depletion of intracellular pyrimidine pools and (ii) the compound may also exert its antiviral activity through inhibition of virion assembly/release.

Original languageEnglish (US)
Pages (from-to)3686-3695
Number of pages10
JournalAntimicrobial Agents and Chemotherapy
Volume54
Issue number9
DOIs
StatePublished - Sep 2010
Externally publishedYes

Fingerprint

brequinar
Dengue Virus
Cell Culture Techniques
Mutation
Viruses
Virion
RNA
Western Equine Encephalitis Viruses
Rhabdoviridae
Yellow fever virus
Alphavirus
Viral Envelope Proteins
Tick-Borne Encephalitis Viruses
Flavivirus
Cytidine
West Nile virus
Pyrimidines
Purines
Vesicular Stomatitis
Proteins

ASJC Scopus subject areas

  • Pharmacology (medical)
  • Pharmacology
  • Infectious Diseases

Cite this

Characterization of dengue virus resistance to brequinar in cell culture. / Qing, Min; Zou, Gang; Wang, Qing Yin; Xu, Hao Ying; Dong, Hongping; Yuan, Zhiming; Shi, Pei-Yong.

In: Antimicrobial Agents and Chemotherapy, Vol. 54, No. 9, 09.2010, p. 3686-3695.

Research output: Contribution to journalArticle

Qing, Min ; Zou, Gang ; Wang, Qing Yin ; Xu, Hao Ying ; Dong, Hongping ; Yuan, Zhiming ; Shi, Pei-Yong. / Characterization of dengue virus resistance to brequinar in cell culture. In: Antimicrobial Agents and Chemotherapy. 2010 ; Vol. 54, No. 9. pp. 3686-3695.
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