In vitro assembly of nucleocapsid-like particles from purified recombinant capsid protein of dengue-2 virus

López, Carlos; Gil, Lázaro; Lazo, Laura; Menéndez, Ivón; Marcos, Ernesto; Sánchez, Jorge; Valdés, Iris; Falcón, Viviana; Rosa, Maria Cristina; Márquez, Gabriel; Guillén, Gerardo; Hermida, Lisset

doi:10.1007/s00705-009-0350-8

Cited by 42 publications

(36 citation statements)

References 19 publications

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“…But there is a problem: the dimers of the Dengue capsid virus are stable in solution so assembly involves combining 90 such dimers into a capsid [37]. This suggests that the primary order parameter at the solidification transition should be an icosahedral spherical harmonic with 90 maxima but plots of the the Y h (l) for l less than or equal to l = 27 do not reveal any such.…”

Section: Cowpea Chlorotic Mottle Virusmentioning

confidence: 99%

Orientational phase transitions and the assembly of viral capsids

et al. 2017

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We present a generalized Landau-Brazovskii free energy for the solidification of chiral molecules on a spherical surface in the context of the assembly of viral shells. We encounter two types of icosahedral solidification transitions. The first type is a conventional first-order phase transition from the uniform to the icosahedral state. It can be described by a single icosahedral spherical harmonic of even l. The chiral pseudo-scalar term in the free energy creates secondary terms with chiral character but it does not affect the thermodynamics of the transition. The second type, associated with icosahedral spherical harmonics with odd l, is anomalous. Pure odd l icosahedral states are unstable but stability is recovered if admixture with the neighboring l + 1 icosahedral spherical harmonic is included, generated by the non-linear terms. This is in conflict with the principle of Landau theory that symmetry-breaking transitions are characterized by only a single irreducible representation of the symmetry group of the uniform phase and we argue that this principle should be removed from Landau theory. The chiral term now directly affects the transition because it lifts the degeneracy between two isomeric mixed-l icosahedral states. A direct transition is possible only over a limited range of parameters. Outside this range, non-icosahedral states intervene. For the important case of capsid assembly dominated by l = 15, the intervening states are found to be based on octahedral symmetry.

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Section: Cowpea Chlorotic Mottle Virusmentioning

confidence: 99%

Orientational phase transitions and the assembly of viral capsids

et al. 2017

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“…2) (Roby et al, 2012). The exact nature of nucleocapsid assembly remains elusive, as the nucleocapsids themselves display no inherent order to their structure (Kiermayr et al, 2004;Kuhn et al, 2002;Ló pez et al, 2009;Zhang et al, 2007b) and a physical interaction of the nucleocapsid with the envelope is not observed. In accordance, the production of empty prM/E particles during virus infection (Lindenbach et al, 2007;Murray et al, 2008) and when these genes are expressed in the absence of C (Allison et al, 1995(Allison et al, , 2003Konishi & Mason, 1993;Lorenz et al, 2003;Wang et al, 2009) suggests that prM/E formation and nucleocapsid assembly are independent processes.…”

Section: Formation Of the Nucleocapsidmentioning

confidence: 99%

“…Determination of the precise structure of the flavivirus nucleocapsid remains elusive despite documentation of assembly of spherical nucleocapsid-like particles in vitro following incubation of recombinant C with nucleic acids (Kiermayr et al, 2004;Ló pez et al, 2009;Teoh et al, 2014), as well as the visualization of purported nucleocapsids adjacent to VP pores in electron micrographs of DENVinfected cells (Welsch et al, 2009). Regardless of the nature of flavivirus nucleocapsids, their assembly appears to be dependent on the association of C with the ER membrane and lipid droplets, presumably to optimally position the protein between the VP pore where nascent genomic RNA exists and the ER sites of prM/E envelope generation (Samsa et al, 2009;Welsch et al, 2009;Westaway et al, 1997).…”

Section: Formation Of the Nucleocapsidmentioning

confidence: 99%

“…However, the efficiency of C dimerization impacts on the overall integrity of the assembled particle in a thermal stability assay (Patkar et al, 2007), suggesting that some interaction between C and prM/E does still occur in the virion. Recent evidence describing the ability of DENV C to act as a histone mimic and cause nucleic acid condensation (Colpitts et al, 2011) supports the hypothesis that the flavivirus nucleocapsid is not an ordered icosahedral shell surrounding the genome but rather the RNA condenses around C dimers.Determination of the precise structure of the flavivirus nucleocapsid remains elusive despite documentation of assembly of spherical nucleocapsid-like particles in vitro following incubation of recombinant C with nucleic acids (Kiermayr et al, 2004;Ló pez et al, 2009;Teoh et al, 2014), as well as the visualization of purported nucleocapsids adjacent to VP pores in electron micrographs of DENVinfected cells (Welsch et al, 2009). Regardless of the nature of flavivirus nucleocapsids, their assembly appears to be dependent on the association of C with the ER membrane and lipid droplets, presumably to optimally position the protein between the VP pore where nascent genomic RNA …”

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Post-translational regulation and modifications of flavivirus structural proteins

Roby

Setoh

Hall

et al. 2015

Journal of General Virology

102

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Flaviviruses are a group of single-stranded, positive-sense RNA viruses that generally circulate between arthropod vectors and susceptible vertebrate hosts, producing significant human and veterinary disease burdens. Intensive research efforts have broadened our scientific understanding of the replication cycles of these viruses and have revealed several elegant and tightly co-ordinated post-translational modifications that regulate the activity of viral proteins. The three structural proteins in particular -capsid (C), pre-membrane (prM) and envelope (E) -are subjected to strict regulatory modifications as they progress from translation through virus particle assembly and egress. The timing of proteolytic cleavage events at the C-prM junction directly influences the degree of genomic RNA packaging into nascent virions. Proteolytic maturation of prM by host furin during Golgi transit facilitates rearrangement of the E proteins at the virion surface, exposing the fusion loop and thus increasing particle infectivity. Specific interactions between the prM and E proteins are also important for particle assembly, as prM acts as a chaperone, facilitating correct conformational folding of E. It is only once prM/E heterodimers form that these proteins can be secreted efficiently. The addition of branched glycans to the prM and E proteins during virion transit also plays a key role in modulating the rate of secretion, pH sensitivity and infectivity of flavivirus particles. The insights gained from research into post-translational regulation of structural proteins are beginning to be applied in the rational design of improved flavivirus vaccine candidates and make attractive targets for the development of novel therapeutics. FlavivirusesThe genus Flavivirus in the family Flaviviridae comprises small, enveloped viruses with non-segmented genomes consisting of single-stranded, positive-sense RNA. These RNA genomes are flanked by 59 and 39 untranslated regions (UTRs) and encode a single polyprotein that is cleaved coand post-translationally to generate between 10 and 11 viral proteins. The 59 UTR contains a type 1 m 7 GpppNm cap structure and the 39 UTR lacks a polyadenylated tail. Structural elements within the UTRs regulate genome replication, translation and host immune responses. Viral proteins are divided between structural proteins, which form the virion, and non-structural proteins, which are involved in genomic replication and modulating host immunity (Fig. 1a) (Lindenbach et al., 2007;Roby et al., 2012).Flaviviruses are maintained predominantly in natural transmission cycles between vertebrate reservoir species (normally mammalian or avian) and haematophagous arthropod vectors (mosquitoes or ticks). Notable exceptions are the viruses that are maintained solely in mosquito hosts (insectspecific flaviviruses) and viruses with no known invertebrate vector (Cook et al., 2012;Mackenzie et al., 2004). As of 2013, the International Committee on Taxonomy of Viruses has classified 53 different viral species as belonging to ...

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“…Packaging of the viral genome within the capsid has been shown to be coupled with RNA replication and is required for the release of infectious flavivirus particles (8,9). The C protein binds nucleic acids promiscuously; how the DENV genome is targeted to sites of capsid assembly remains an area of active research (10,11). Although the amino terminus of the C protein is not required for protein integrity (6,12), deletions in the region can result in defects in particle formation in human cells (13).…”

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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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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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In vitro assembly of nucleocapsid-like particles from purified recombinant capsid protein of dengue-2 virus

Cited by 42 publications

References 19 publications

Orientational phase transitions and the assembly of viral capsids

Orientational phase transitions and the assembly of viral capsids

Post-translational regulation and modifications of flavivirus structural proteins

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

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