Single point mutation in tick-borne encephalitis virus prM protein induces a reduction of virus particle secretion

Yoshii, Kentarou; Konno, Akihiro; Goto, Akiko; Nio, Junko; Obara, Mayumi; Ueki, Tomotaka; Hayasaka, Daisuke; Mizutani, Tetsuya; Kariwa, Hiroaki; Takashima, Ikuo

doi:10.1099/vir.0.80169-0

Cited by 46 publications

(31 citation statements)

References 52 publications

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“…It is currently believed that flavivirus genomes are released into and replicate in the cytoplasm in close association with intracellular membranous structures that possibly derive from the ER (Clyde et al, 2006;Miller & Krijnse-Locker, 2008). Consistently, flavivirus infections characteristically result in significant proliferation of rough ER membranes, and the flavivirus replication complex has been partly correlated with these ER membranes (Boulton & Westaway, 1976) and with cytoplasmic vesicles and vacuoles (Mackenzie et al, 1996), and it has been proposed that flaviviruses bud from ER membranes and transit the Golgi body before release from the cell (Clyde et al, 2006;Yoshii et al, 2004).…”

Section: Location Of Dengue Virus Replication Complexmentioning

confidence: 71%

“…Consistently, flavivirus infections characteristically result in significant proliferation of rough ER membranes, and the flavivirus replication complex has been partly correlated with these ER membranes (Boulton & Westaway, 1976) and with cytoplasmic vesicles and vacuoles (Mackenzie et al, 1996), and it has been proposed that flaviviruses bud from ER membranes and transit the Golgi body before release from the cell (Clyde et al, 2006;Yoshii et al, 2004).…”

Section: Discussionmentioning

confidence: 82%

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Co-localization of constituents of the dengue virus translation and replication machinery with amphisomes

Panyasrivanit

Khakpoor

Wikan

et al. 2009

Journal of General Virology

148

176

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Infections with dengue virus (DENV) are a significant public health concern in tropical and subtropical regions. However, little detail is known about how DENV interacts with the host-cell machinery to facilitate its translation and replication. In DENV-infected HepG2 cells, an increase in the level of LC3-II (microtubule-associated protein 1 light chain 3 form II), the autophagosomal membrane-bound form of LC3, was observed, and LC3 was found to co-localize with dsRNA and DENV NS1 protein, as well as ribosomal protein L28, indicating the presence of at least some of the DENV translation/replication machinery on autophagic vacuoles. Inhibition of fusion of autophagic vacuoles with lysosomes resulted in an increase in both intracellular and extracellular virus, and co-localization observed between mannose-6-phosphate receptor (MPR) and dsRNA and between MPR and LC3 identified the autophagic vacuoles as amphisomes. Amphisomes are formed as a result of fusion between endosomal and autophagic vacuoles, and as such provide a direct link between virus entry and subsequent replication and translation. INTRODUCTIONWith an estimated 100 million infections per year worldwide, dengue virus (DENV), which is spread to humans by the bite of female Aedes mosquitoes, represents a significant public health threat in tropical and subtropical countries (Guzman & Kouri, 2002). The DENV complex comprises four antigenically distinct viruses termed DENV serotypes 1, 2, 3 and 4 (DENV-1 to -4), all of which can cause a wide spectrum of disease presentation, from a relatively mild febrile disease to a life-threatening haemorrhagic syndrome (Malavige et al., 2004). DENV is an enveloped, positivesense, single-stranded RNA virus of approximately 11 kb that encodes three structural proteins (core, pre-membrane and envelope) and seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5) in one open reading frame (Chang, 1997). Entry of DENV into a susceptible cell occurs primarily through receptormediated endocytosis via clathrin-coated pits (Krishnan et al., 2007) and the virus is then trafficked to endosomes (Krishnan et al., 2007;van der Schaar et al., 2007) and fuses with the endosomal membranes (Heinz et al., 2004) through a pH-dependent conformational change in the envelope protein (Modis et al., 2004;Mukhopadhyay et al., 2005). The fate of the released nucleocapsid is largely obscure, although it is known that, after uncoating of the virus genome, the host-cell translational machinery is utilized to synthesize a polyprotein precursor that is processed co-and post-translationally by a virus-encoded protease (NS3) and host signallases (Cahour et al., 1992). Translation and subsequent replication of the DENV genome occur in tight association with intracellular membranous structures that are believed to be endoplasmic reticulum (ER)-derived (Clyde et al., 2006;Miller & Krijnse-Locker, 2008;Salonen et al., 2005), although the exact origin of these membranes remains unclear.Autophagy is a lysosomal degradation pathway involve...

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Section: Location Of Dengue Virus Replication Complexmentioning

confidence: 71%

Section: Discussionmentioning

confidence: 82%

Co-localization of constituents of the dengue virus translation and replication machinery with amphisomes

Panyasrivanit

Khakpoor

Wikan

et al. 2009

Journal of General Virology

148

176

View full text Add to dashboard Cite

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“…DENV2 protein production in MEP cell lines appeared to be coordinated, leading to lower amounts of excessive viral protein production, thereby reducing the likelihood of immune recognition. Many mutations associated with reduced virus production have already been identified (Junjhon et al, 2008; Lee et al, 2010; Pryor et al, 2004; Yoshii et al, 2004) and could potentially play a role in shaping virus particle production in MEP cells.…”

Section: Discussionmentioning

confidence: 99%

Characterization of dengue virus 2 growth in megakaryocyte–erythrocyte progenitor cells

et al. 2016

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Megakaryocyte–erythrocyte progenitor (MEP) cells are potential in vivo targets of dengue virus (DENV); the virus has been found associated with megakaryocytes ex vivo and platelets during DENV-induced thrombocytopenia. We report here that DENV serotype 2 (DENV2) propagates well in human non-differentiated MEP cell lines (Meg01 and K562). In comparison to virus propagated in Vero cells, viruses from MEP cell lines had similar structure and buoyant density. However, differences in MEP-DENV2 stability and composition were suggested by distinct protein patterns in western blot analysis. Also, antibody neutralization of envelope domain I/II on MEP-DENV2 was reduced relative to that on Vero-DENV2. Infectious DENV2 was produced at comparable kinetics and magnitude in MEP and Vero cells. However, fewer virion structures appeared in electron micrographs of MEP cells. We propose that DENV2 infects and produces virus efficiently in megakaryocytes and that megakaryocyte impairment might contribute to dengue disease pathogenesis.

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“…These mutations may affect the particles being formed in the ER if they are associated with a structural change that favors one particle type over another. Certain prM mutations are known to affect the assembly or stability of the viral particles (34,51). Recent evidence revealed the proprotein convertase activity in the ER and early Golgi apparatus compartments (41), suggesting that the pr-M junction mutations may affect immature particle-furin interaction soon after budding.…”

Section: Discussionmentioning

confidence: 99%

Differential Modulation of prM Cleavage, Extracellular Particle Distribution, and Virus Infectivity by Conserved Residues at Nonfurin Consensus Positions of the Dengue Virus pr-M Junction

Junjhon

Lausumpao

Supasa

et al. 2008

J Virol

110

137

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In the generation of flavivirus particles, an internal cleavage of the envelope glycoprotein prM by furin is required for the acquisition of infectivity. Unlike cleavage of the prM of other flaviviruses, cleavage of dengue virus prM is incomplete in many cell lines; the partial cleavage reflects the influence of residues at furin nonconsensus positions of the pr-M junction, as flaviviruses share basic residues at positions P1, P2, and P4, recognized by furin. In this study, viruses harboring the alanine-scanning and other multiple-point mutations of the pr-M junction were generated, employing a dengue virus background that exhibited 60 to 70% prM cleavage and a preponderance of virion-sized extracellular particles. Analysis of prM and its cleavage products in viable mutants revealed a cleavage-suppressive effect at the conserved P3 Glu residue, as well as the cleavage-augmenting effects at the P5 Arg and P6 His residues, indicating an interplay between opposing modulatory influences mediated by these residues on the cleavage of the pr-M junction. Changes in the prM cleavage level were associated with altered proportions of extracellular virions and subviral particles; mutants with reduced cleavage were enriched with subviral particles and prM-containing virions, whereas the mutant with enhanced cleavage was deprived of these particles. Alterations of virus multiplication were detected in mutants with reduced prM cleavage and were correlated with their low specific infectivities. These findings define the functional roles of charged residues located adjacent to the furin consensus sequence in the cleavage of dengue virus prM and provide plausible mechanisms by which the reduction in the pr-M junction cleavability may affect virus replication.Dengue viruses are members of the genus Flavivirus in the family Flaviviridae. A single-stranded genomic RNA of positive polarity encodes an open reading frame which is translated into a precursor of three structural proteins and at least seven nonstructural proteins (27). During the replication of flaviviruses, proteolytic cleavages of the polyprotein by viral protease and a number of cellular enzymes occur in distinct subcellular compartments (27,32). Three structural proteins, C, prM/M, and E, are associated with the membrane of the rough endoplasmic reticulum (ER) by C-terminal membrane-spanning regions. Cleavages distal to the membrane-spanning regions of C and the two envelope glycoproteins prM and E by host signalase in the lumen of the rough ER and a cleavage proximal to the membrane-spanning region of C by viral protease in the cytoplasm are important to the assembly of viral particles (32). During the export of immature particles through the secretory pathway, cleavage of prM by the trans-Golgi apparatus resident furin allows reorganization of the receptor-binding E protein that is required for the acquisition of infectivity (9, 43).Assembly of flavivirus particles in the rough ER results in two types of particles: virions and subviral particles (32,40). An immature...

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Single point mutation in tick-borne encephalitis virus prM protein induces a reduction of virus particle secretion

Cited by 46 publications

References 52 publications

Co-localization of constituents of the dengue virus translation and replication machinery with amphisomes

Co-localization of constituents of the dengue virus translation and replication machinery with amphisomes

Characterization of dengue virus 2 growth in megakaryocyte–erythrocyte progenitor cells

Differential Modulation of prM Cleavage, Extracellular Particle Distribution, and Virus Infectivity by Conserved Residues at Nonfurin Consensus Positions of the Dengue Virus pr-M Junction

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