Understanding Dengue Virus Capsid Protein Disordered N-Terminus and pep14-23-Based Inhibition

Faustino, André F.; Guerra, Grazia Maria; Huber, Roland G.; Hollmann, Axel; Domingues, Marco M.; Barbosa, G.M.; Enguita, Francisco J.; Bond, Peter J.; Castanho, Miguel A. R. B.; Poian, Andrea T. Da; Almeida, Fábio C. L.; Santos, Nuno C.; Martins, Ivo C.

doi:10.1021/cb500640t

Cited by 51 publications

(85 citation statements)

References 53 publications

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“…The charge calculated for NSP1-CTR was -6, which thus cause electrostatic repulsion while interacting with DOPS. Ionic interactions play important role in interaction of peptide with oppositely charged lipid vesicals as reported previously (36,53).…”

Section: Nsp1-ctr Conformational Change Revealed By Trp Fluorescencesupporting

confidence: 52%

SARS-CoV-2 NSP1 C-terminal region (residues 130-180) is an intrinsically disordered region

Kumar¹,

Kumar²,

Kumar³

et al. 2020

Preprint

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Nonstructural protein 1 (NSP1) of SARS-CoV-2 plays a key role in downregulation of RIG-I pathways and interacts with 40 S ribosome. Recently, the cryo-EM structure in complex with 40S ribosome is deciphered. However, the structure of full length NSP1 without any partner has not been studies. Also, the conformation of NSP1-C terminal region in isolation is not been studied. In this study, we have investigated the conformational dynamics of NSP1C-terminal region (NSP1-CTR; amino acids 130-180) in isolation and under different solvent environments. The NSP1-CTR is found to be intrinsically disordered in aqueous solution. Further, we used alpha helix inducer, trifluoroethanol, and found induction of alpha helical conformation using CD spectroscopy. Additionally, in the presence of SDS, NSP1-CTR is showing a conformational change from disordered to ordered, possibly gaining alpha helix in part. But in presence of neutral lipid DOPC, a slight change in conformation is observed. This implies the possible role of hydrophobic interaction and electrostatic interaction on the conformational changes of NSP1. The changes in structural conformation were further studied by fluorescence-based studies, which showed significant blue shift and fluorescence quenching in the presence of SDS and TFE. Lipid vesicles also showed fluorescence-based quenching. In agreement to these result, fluorescence lifetime and fluorescence anisotropy decay suggests a change in conformational dynamics. The zeta potential studies further validated that the conformational dynamics is mostly because of hydrophobic interaction. In last, these experimental studies were complemented through Molecular Dynamics (MD) simulation which have also shown a good correlation and testify our experiments. We believe that the intrinsically disordered nature of the NSP1-CTR will have implications in disorder based binding promiscuity with its interacting proteins.

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Section: Nsp1-ctr Conformational Change Revealed By Trp Fluorescencesupporting

confidence: 52%

SARS-CoV-2 NSP1 C-terminal region (residues 130-180) is an intrinsically disordered region

Kumar¹,

Kumar²,

Kumar³

et al. 2020

Preprint

View full text Add to dashboard Cite

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“…In vitro studies using atomic force microscopy provided evidence that a peptide corresponding to the disordered N-terminal region of capsid interacts with negatively charged LDs, suggesting that this region also facilitates LD binding (82). Further studies indicated that the N-terminal peptide inhibits, in a dose-dependent manner, in vitro binding of capsid to LDs (83). It has also been proposed that capsid binding to LDs depends on high concentrations of potassium and that this binding could be mediated by the LD-associated protein TIP47 (84).…”

Section: Subcellular Distribution Of Capsidmentioning

confidence: 99%

“…A DENV capsid inhibitor based on a peptide mimic of the N-terminal region of the viral protein has been proposed (82, 83). This peptide has been suggested to compete with different capsid functions, such as LD association.…”

Section: Capsid Protein and Antiviral Strategiesmentioning

confidence: 99%

Properties and Functions of the Dengue Virus Capsid Protein

2016

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Dengue virus affects hundreds of millions of people each year around the world, causing a tremendous social and economic impact on affected countries. The aim of this review is to summarize our current knowledge of the functions, structure, and interactions of the viral capsid protein. The primary role of capsid is to package the viral genome. There are two processes linked to this function: the recruitment of the viral RNA during assembly and the release of the genome during infection. Although particle assembly takes place on endoplasmic reticulum membranes, capsid localizes in nucleoli and lipid droplets. Why capsid accumulates in these locations during infection remains unknown. In this review, we describe available data and discuss new ideas on dengue virus capsid functions and interactions. We believe that a deeper understanding of how the capsid protein works during infection will create opportunities for novel antiviral strategies, which are urgently needed to control dengue virus infections. KeywordsRNA virus; flavivirus; arbovirus; dengue virus; capsid protein; viral encapsidation; viral assembly; lipid droplets DENGUE VIRUSDengue virus (DENV) is the most significant arthropod-borne viral pathogen in humans. The geographical spread and incidence of DENV infections have increased dramatically in recent years. DENV is estimated to cause around 390 million infections per year, placing over 3 billion people at risk of infection (1). In addition to the heavy burden placed on public health, DENV epidemics have a huge economic impact on affected countries.DENV is a member of the Flavivirus genus of the Flaviviridae family (2). The Flavivirus genus includes other important emerging and reemerging human pathogens such as Zika virus (ZIKV), West Nile virus (WNV), Japanese encephalitis virus ( JEV), yellow fever virus (YFV), and Saint Louis encephalitis virus (SLEV) (3). Most flaviviruses are arthropodborne; however, vertebrate-and invertebrate-specific viruses are also members of the group (for a recent review see 4). DENV cycles in nature between Aedes mosquito vectors (mainly DISCLOSURE STATEMENTThe authors are not aware of any affiliations, memberships, funding, or financial holdings that might be perceived as affecting the objectivity of this review. HHS Public Access Author Manuscript Author ManuscriptAuthor Manuscript Author ManuscriptAedes albopictus and Aedes aegypti ) and humans. Four DENV serotypes (DENV1, DENV2, DENV3, and DENV4) circulate in tropical and subtropical regions of the globe (5). They differ from one another by 25-40% at the amino acid level and are further separated into genotypes. Clinical outcomes for all serotypes can be unapparent or result in a spectrum of diseases ranging from self-limited dengue fever to severe dengue, a potentially lethal hemorrhagic illness. The incidence of dengue disease is growing as the mosquito vector spreads owing to urbanization, population growth, international travel, insufficient mosquito control efforts, and global warming.Although vaccines a...

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“…11 It was also used for understanding the molecular determinants and identifying the ligand for the dengue virus capsid protein on intracellular lipid droplets and plasma lipoproteins (Figure 4 .3) . 17,[47][48][49][50] Glass surface…”

Section: Single-molecule Interactionsmentioning

confidence: 99%