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Virus-Cell Interactions

Cross-Inhibition of Chikungunya Virus Fusion and Infection by Alphavirus E1 Domain III Proteins

Claudia Sánchez-San Martín, Soumya Nanda, Yan Zheng, Whitney Fields, Margaret Kielian
Claudia Sánchez-San Martín
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA
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Soumya Nanda
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA
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Yan Zheng
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA
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Whitney Fields
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA
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Margaret Kielian
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA
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DOI: 10.1128/JVI.00814-13
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ABSTRACT

Alphaviruses are small enveloped RNA viruses that include important emerging human pathogens, such as chikungunya virus (CHIKV). These viruses infect cells via a low-pH-triggered membrane fusion reaction, making this step a potential target for antiviral therapies. The E1 fusion protein inserts into the target membrane, trimerizes, and refolds to a hairpin-like conformation in which the combination of E1 domain III (DIII) and the stem region (DIII-stem) pack against a core trimer composed of E1 domains I and II (DI/II). Addition of exogenous DIII proteins from Semliki Forest virus (SFV) has been shown to inhibit E1 hairpin formation and SFV fusion and infection. Here we produced and characterized DIII and DI/II proteins from CHIKV and SFV. Unlike SFV DIII, both core trimer binding and fusion inhibition by CHIKV DIII required the stem region. CHIKV DIII-stem and SFV DIII-stem showed efficient cross-inhibition of SFV, Sindbis virus, and CHIKV infections. We developed a fluorescence anisotropy-based assay for the binding of SFV DIII-stem to the core trimer and used it to demonstrate the relatively high affinity of this interaction (Kd [dissociation constant], ∼85 nM) and the importance of the stem region. Together, our results support the conserved nature of the key contacts of DIII-stem in the alphavirus E1 homotrimer and describe a sensitive and quantitative in vitro assay for this step in fusion protein refolding.

  • Copyright © 2013, American Society for Microbiology. All Rights Reserved.
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Cross-Inhibition of Chikungunya Virus Fusion and Infection by Alphavirus E1 Domain III Proteins
Claudia Sánchez-San Martín, Soumya Nanda, Yan Zheng, Whitney Fields, Margaret Kielian
Journal of Virology Jun 2013, 87 (13) 7680-7687; DOI: 10.1128/JVI.00814-13

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Cross-Inhibition of Chikungunya Virus Fusion and Infection by Alphavirus E1 Domain III Proteins
Claudia Sánchez-San Martín, Soumya Nanda, Yan Zheng, Whitney Fields, Margaret Kielian
Journal of Virology Jun 2013, 87 (13) 7680-7687; DOI: 10.1128/JVI.00814-13
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