Inhibition of dengue virus by targeting viral NS4B protein

Xuping Xie, Qing Yin Wang, Hao Ying Xu, Min Qing, Laura Kramer, Zhiming Yuan, Pei-Yong Shi

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

We report a novel inhibitor that selectively suppresses dengue virus (DENV) by targeting viral NS4B protein. The inhibitor was identified by screening a 1.8-million-compound library using a luciferase replicon of DENV serotype 2 (DENV-2). The compound specifically inhibits all four serotypes of DENV (50% effective concentration [EC 50], 1 to 4 μM; and 50% cytotoxic concentration [CC 50], >40 μM), but it does not inhibit closely related flaviviruses (West Nile virus and yellow fever virus) or nonflaviviruses (Western equine encephalomyelitis virus, Chikungunya virus, and vesicular stomatitis virus). A mode-of-action study suggested that the compound inhibits viral RNA synthesis. Replicons resistant to the inhibitor were selected in cell culture. Sequencing of the resistant replicons revealed two mutations (P104L and A119T) in the viral NS4B protein. Genetic analysis, using DENV-2 replicon and recombinant viruses, demonstrated that each of the two NS4B mutations alone confers partial resistance and double mutations confer additive resistance to the inhibitor in mammalian cells. In addition, we found that a replication defect caused by a lethal NS4B mutation could be partially rescued through trans complementation. The ability to complement NS4B in trans affected drug sensitivity when a single cell was coinfected with drug-sensitive and drug-resistant viruses. Mechanistically, NS4B was previously shown to interact with the viral NS3 helicase domain; one of the two NS4B mutations recovered in our resistance analysis-P104L-abolished the NS3-NS4B interaction (I. Umareddy, A. Chao, A. Sampath, F. Gu, and S. G. Vasudevan, J. Gen. Virol. 87:2605-2614, 2006). Collectively, the results suggest that the identified inhibitor targets the DENV NS4B protein, leading to a defect in viral RNA synthesis.

Original languageEnglish (US)
Pages (from-to)11183-11195
Number of pages13
JournalJournal of Virology
Volume85
Issue number21
DOIs
StatePublished - Nov 2011
Externally publishedYes

Fingerprint

Dengue virus
Dengue Virus
viral proteins
Viral Proteins
replicon
Replicon
mutation
Mutation
Viral RNA
Viruses
drugs
West Nile Fever
Western Equine Encephalitis Viruses
Yellow fever virus
Western equine encephalitis virus
Chikungunya virus
Pharmaceutical Preparations
RNA
Flaviviridae
Vesiculovirus

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Xie, X., Wang, Q. Y., Xu, H. Y., Qing, M., Kramer, L., Yuan, Z., & Shi, P-Y. (2011). Inhibition of dengue virus by targeting viral NS4B protein. Journal of Virology, 85(21), 11183-11195. https://doi.org/10.1128/JVI.05468-11

Inhibition of dengue virus by targeting viral NS4B protein. / Xie, Xuping; Wang, Qing Yin; Xu, Hao Ying; Qing, Min; Kramer, Laura; Yuan, Zhiming; Shi, Pei-Yong.

In: Journal of Virology, Vol. 85, No. 21, 11.2011, p. 11183-11195.

Research output: Contribution to journalArticle

Xie, X, Wang, QY, Xu, HY, Qing, M, Kramer, L, Yuan, Z & Shi, P-Y 2011, 'Inhibition of dengue virus by targeting viral NS4B protein', Journal of Virology, vol. 85, no. 21, pp. 11183-11195. https://doi.org/10.1128/JVI.05468-11
Xie X, Wang QY, Xu HY, Qing M, Kramer L, Yuan Z et al. Inhibition of dengue virus by targeting viral NS4B protein. Journal of Virology. 2011 Nov;85(21):11183-11195. https://doi.org/10.1128/JVI.05468-11
Xie, Xuping ; Wang, Qing Yin ; Xu, Hao Ying ; Qing, Min ; Kramer, Laura ; Yuan, Zhiming ; Shi, Pei-Yong. / Inhibition of dengue virus by targeting viral NS4B protein. In: Journal of Virology. 2011 ; Vol. 85, No. 21. pp. 11183-11195.
@article{218c5e94b1b543ff89fb677f3c5fc23e,
title = "Inhibition of dengue virus by targeting viral NS4B protein",
abstract = "We report a novel inhibitor that selectively suppresses dengue virus (DENV) by targeting viral NS4B protein. The inhibitor was identified by screening a 1.8-million-compound library using a luciferase replicon of DENV serotype 2 (DENV-2). The compound specifically inhibits all four serotypes of DENV (50{\%} effective concentration [EC 50], 1 to 4 μM; and 50{\%} cytotoxic concentration [CC 50], >40 μM), but it does not inhibit closely related flaviviruses (West Nile virus and yellow fever virus) or nonflaviviruses (Western equine encephalomyelitis virus, Chikungunya virus, and vesicular stomatitis virus). A mode-of-action study suggested that the compound inhibits viral RNA synthesis. Replicons resistant to the inhibitor were selected in cell culture. Sequencing of the resistant replicons revealed two mutations (P104L and A119T) in the viral NS4B protein. Genetic analysis, using DENV-2 replicon and recombinant viruses, demonstrated that each of the two NS4B mutations alone confers partial resistance and double mutations confer additive resistance to the inhibitor in mammalian cells. In addition, we found that a replication defect caused by a lethal NS4B mutation could be partially rescued through trans complementation. The ability to complement NS4B in trans affected drug sensitivity when a single cell was coinfected with drug-sensitive and drug-resistant viruses. Mechanistically, NS4B was previously shown to interact with the viral NS3 helicase domain; one of the two NS4B mutations recovered in our resistance analysis-P104L-abolished the NS3-NS4B interaction (I. Umareddy, A. Chao, A. Sampath, F. Gu, and S. G. Vasudevan, J. Gen. Virol. 87:2605-2614, 2006). Collectively, the results suggest that the identified inhibitor targets the DENV NS4B protein, leading to a defect in viral RNA synthesis.",
author = "Xuping Xie and Wang, {Qing Yin} and Xu, {Hao Ying} and Min Qing and Laura Kramer and Zhiming Yuan and Pei-Yong Shi",
year = "2011",
month = "11",
doi = "10.1128/JVI.05468-11",
language = "English (US)",
volume = "85",
pages = "11183--11195",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "21",

}

TY - JOUR

T1 - Inhibition of dengue virus by targeting viral NS4B protein

AU - Xie, Xuping

AU - Wang, Qing Yin

AU - Xu, Hao Ying

AU - Qing, Min

AU - Kramer, Laura

AU - Yuan, Zhiming

AU - Shi, Pei-Yong

PY - 2011/11

Y1 - 2011/11

N2 - We report a novel inhibitor that selectively suppresses dengue virus (DENV) by targeting viral NS4B protein. The inhibitor was identified by screening a 1.8-million-compound library using a luciferase replicon of DENV serotype 2 (DENV-2). The compound specifically inhibits all four serotypes of DENV (50% effective concentration [EC 50], 1 to 4 μM; and 50% cytotoxic concentration [CC 50], >40 μM), but it does not inhibit closely related flaviviruses (West Nile virus and yellow fever virus) or nonflaviviruses (Western equine encephalomyelitis virus, Chikungunya virus, and vesicular stomatitis virus). A mode-of-action study suggested that the compound inhibits viral RNA synthesis. Replicons resistant to the inhibitor were selected in cell culture. Sequencing of the resistant replicons revealed two mutations (P104L and A119T) in the viral NS4B protein. Genetic analysis, using DENV-2 replicon and recombinant viruses, demonstrated that each of the two NS4B mutations alone confers partial resistance and double mutations confer additive resistance to the inhibitor in mammalian cells. In addition, we found that a replication defect caused by a lethal NS4B mutation could be partially rescued through trans complementation. The ability to complement NS4B in trans affected drug sensitivity when a single cell was coinfected with drug-sensitive and drug-resistant viruses. Mechanistically, NS4B was previously shown to interact with the viral NS3 helicase domain; one of the two NS4B mutations recovered in our resistance analysis-P104L-abolished the NS3-NS4B interaction (I. Umareddy, A. Chao, A. Sampath, F. Gu, and S. G. Vasudevan, J. Gen. Virol. 87:2605-2614, 2006). Collectively, the results suggest that the identified inhibitor targets the DENV NS4B protein, leading to a defect in viral RNA synthesis.

AB - We report a novel inhibitor that selectively suppresses dengue virus (DENV) by targeting viral NS4B protein. The inhibitor was identified by screening a 1.8-million-compound library using a luciferase replicon of DENV serotype 2 (DENV-2). The compound specifically inhibits all four serotypes of DENV (50% effective concentration [EC 50], 1 to 4 μM; and 50% cytotoxic concentration [CC 50], >40 μM), but it does not inhibit closely related flaviviruses (West Nile virus and yellow fever virus) or nonflaviviruses (Western equine encephalomyelitis virus, Chikungunya virus, and vesicular stomatitis virus). A mode-of-action study suggested that the compound inhibits viral RNA synthesis. Replicons resistant to the inhibitor were selected in cell culture. Sequencing of the resistant replicons revealed two mutations (P104L and A119T) in the viral NS4B protein. Genetic analysis, using DENV-2 replicon and recombinant viruses, demonstrated that each of the two NS4B mutations alone confers partial resistance and double mutations confer additive resistance to the inhibitor in mammalian cells. In addition, we found that a replication defect caused by a lethal NS4B mutation could be partially rescued through trans complementation. The ability to complement NS4B in trans affected drug sensitivity when a single cell was coinfected with drug-sensitive and drug-resistant viruses. Mechanistically, NS4B was previously shown to interact with the viral NS3 helicase domain; one of the two NS4B mutations recovered in our resistance analysis-P104L-abolished the NS3-NS4B interaction (I. Umareddy, A. Chao, A. Sampath, F. Gu, and S. G. Vasudevan, J. Gen. Virol. 87:2605-2614, 2006). Collectively, the results suggest that the identified inhibitor targets the DENV NS4B protein, leading to a defect in viral RNA synthesis.

UR - http://www.scopus.com/inward/record.url?scp=80055106240&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80055106240&partnerID=8YFLogxK

U2 - 10.1128/JVI.05468-11

DO - 10.1128/JVI.05468-11

M3 - Article

C2 - 21865382

AN - SCOPUS:80055106240

VL - 85

SP - 11183

EP - 11195

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 21

ER -