RNA binding property and RNA chaperone activity of dengue virus core protein and other viral RNA-interacting proteins

Pong, Wen-Li; Huang, Zhi-Shun; Teoh, Pak-Guan; Wang, Chung-Chun; Wu, Hurng‐Sheng

doi:10.1016/j.febslet.2011.06.038

Cited by 53 publications

(56 citation statements)

References 23 publications

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“…Both DENV C and NCL can bind RNA (25,34). In order to determine if interactions between DENV C and NCL were occurring through RNA intermediates, co-IP experiments were performed using RNase A-treated lysates from cells transfected with the GFP-DVC expression vector.…”

Section: Resultsmentioning

confidence: 99%

“…Within the cytoplasm, the DENV C protein not only is localized to sites of virus assembly, but is found in association with lipid droplets, where it may play an important role in the formation of virus particles (18)(19)(20)(21). The DENV C protein is also localized in the nuclei of infected cells, where it is primarily associated with the nucleolus (22)(23)(24)(25). The role of the C protein in the nucleus has not yet been elucidated.…”

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confidence: 99%

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Nucleolin Interacts with the Dengue Virus Capsid Protein and Plays a Role in Formation of Infectious Virus Particles

Balinsky

Schmeisser

Ganesan

et al. 2013

J Virol

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Dengue virus (DENV), a member of the genus Flavivirus, causes a spectrum of disease in humans that can range from a mild febrile illness to potentially fatal hemorrhage or shock syndromes. Four serotypes of DENV (DENV-1, -2, -3, and -4) are transmitted by the bite of a mosquito vector and are responsible for more than 50 million infections each year. Infection with one DENV serotype does not confer lasting immunity to the others (1-3). The most severe clinical manifestations of dengue are associated with secondary infections by a heterologous DENV serotype (2). Increasing urbanization, cocirculation of multiple DENV serotypes within the same geographic area, and expansion of its insect vector contribute to the increasing importance of dengue to global health (2, 3).DENV contains a positive-sense single-stranded RNA (ssRNA) genome that is translated as a single open reading frame. The resulting polyprotein is cleaved by virus and host proteases into three structural and at least seven nonstructural proteins (4, 5). The capsid (C) protein functions as a structural component of the DENV virion, encapsulating the ssRNA genome of the virus (4, 5). The DENV C protein is composed of four alpha helices with an unstructured amino terminus and forms antiparallel homodimers.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Nucleolin Interacts with the Dengue Virus Capsid Protein and Plays a Role in Formation of Infectious Virus Particles

Balinsky

Schmeisser

Ganesan

et al. 2013

J Virol

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“…For the cross-linking study, recombinant CP was purified as described previously (15). For cross-linking of purified protein, a volume of 10 l containing the purified CP dissolved in 50 mM HEPES, pH 7.9, 0.1 M NaCl, 1 mM EDTA, and 2% glycerol was preincubated for 15 min at 25°C.…”

Section: Cells and Virusmentioning

confidence: 99%

“…For recombinant CP purification, a plasmid expressing the mature form of DENV-3 (strain Philippines/H87/1956) CP was constructed in a modified pET21b vector in which a stop codon was inserted upstream of the His tag coding sequence (15). Vectors for the expression of CP mutants were generated using site-directed mutagenesis (SDM) ( Table 1).…”

Section: Cells and Virusmentioning

confidence: 99%

“…The dimer conformation and surface positive charge distribution of DENV CP suggest that nucleocapsid assembly occurs through lipid membrane interaction on the ␣2-␣2= hydrophobic surface and genome RNA interaction on the other positively charged surface, the ␣4-␣4= surface (11). CP interacts with RNA nonspecifically, and the RNA chaperone activity displayed by flavivirus CP may play an important role in assisting nucleocapsid formation (14,15).…”

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Maintenance of Dimer Conformation by the Dengue Virus Core Protein α4-α4′ Helix Pair Is Critical for Nucleocapsid Formation and Virus Production

Teoh

Huang

Pong

et al. 2014

J Virol

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The virion of dengue virus (DENV) is composed of a viral envelope covering a nucleocapsid formed by a complex of viral genomic RNA and core protein (CP). DENV CP forms a dimer via the internal ␣2 and ␣4 helices of each monomer. Pairing of ␣2-␣2= creates a continuous hydrophobic surface, while the ␣4-␣4= helix pair joins the homodimer via side-chain interactions of the inner-edge residues. However, the importance of dimer conformation and the ␣4 helix of DENV CP in relation to its function are poorly understood. Loss of association between CP and lipid droplets (LDs) due to mutation suggests that the CP hydrophobic surface was not exposed, offering a possible explanation for the absence of dimers. Further assays suggest the connection between CP folding and protein stability. Attenuation of full-length RNA-derived virus production is associated with CP mutation, since no significant defects were detected in virus translation and replication. The in vitro characterization assays further highlighted that the ␣4-␣4= helix pair conformation is critical in preserving the overall ␣-helical content, thermostability, and dimer formation ability of CP, features correlated with the efficiency of nucleocapsid formation. Addition of Tween 20 improves in vitro nucleocapsid-like particle formation, suggesting the role of the LD in nucleocapsid formation in vivo. This study provides the first direct link between the ␣4-␣4= helix pair interaction and the CP dimer conformation that is the basis of CP function, particularly in nucleocapsid formation during virion production. IMPORTANCEStructure-based mutagenesis study of the dengue virus core protein (CP) reveals that the ␣4-␣4= helix pair is the key to maintaining its dimer conformation, which is the basis of CP function in nucleocapsid formation and virus production. Attenuation of full-length RNA-derived virus production is associated with CP mutation, since no significant defects in virus translation and replication were detected. In vitro inefficiency and size of nucleocapsid-like particle (NLP) formation offer a possible explanation for in vivo virus production inefficiency upon CP mutation. Further, the transition of NLP morphology from an incomplete state to an intact particle shown by ␣4-␣4= helix pair mutants in the presence of a nonionic detergent suggests the regulatory role of the intracellular lipid droplet (LD) in CP-LD interaction and in promoting nucleocapsid formation. This study provides the first direct link between the ␣4-␣4= helix pair interaction and CP dimer conformation that is the fundamental requirement of CP function, particularly in nucleocapsid formation during virion production. V iruses of the Flaviviridae family are enveloped and have a monopartite, linear, and positive-polarity single-stranded RNA genome. A member of this family in the Flavivirus genus, Dengue virus (DENV), is the most important human arthropodborne viral pathogen, putting 3 billion people at risk and resulting in 50 million to 100 million infections and around 22,000 deat...

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Revisiting Dengue Virus–Host Cell Interaction

Acosta

Kumar

Bartenschlager

2014

Advances in Virus Research

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RNA binding property and RNA chaperone activity of dengue virus core protein and other viral RNA-interacting proteins

Cited by 53 publications

References 23 publications

Nucleolin Interacts with the Dengue Virus Capsid Protein and Plays a Role in Formation of Infectious Virus Particles

Nucleolin Interacts with the Dengue Virus Capsid Protein and Plays a Role in Formation of Infectious Virus Particles

Maintenance of Dimer Conformation by the Dengue Virus Core Protein α4-α4′ Helix Pair Is Critical for Nucleocapsid Formation and Virus Production

Revisiting Dengue Virus–Host Cell Interaction

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