Different Architectures in the Assembly of Infectious Bursal Disease Virus Capsid Proteins Expressed in Insect Cells
Identifieur interne : 003748 ( Main/Exploration ); précédent : 003747; suivant : 003749Different Architectures in the Assembly of Infectious Bursal Disease Virus Capsid Proteins Expressed in Insect Cells
Auteurs : Jorge Luis Martinez-Torrecuadrada [Espagne] ; José R. Cast N [Espagne] ; Marian Castro [Espagne] ; José L. Carrascosa [Espagne] ; José F. Rodriguez [Espagne] ; J. Ignacio Casal [Espagne]Source :
- Virology [ 0042-6822 ] ; 2000.
English descriptors
- Teeft :
- Antigenic properties, Antiserum, Assembly products, Baculovirus, Baculovirus transfer vectors, Baculovirus vectors, Bluetongue virus, Bursal, Capsid, Capsid protein, Capsomere lattice, Carrascosa, Cellular extracts, Confocal, Confocal microscopy, Different architectures, Different baculovirus transfer vectors, Different types, Electron microscopy, Electron microscopy analysis, Escherichia coli, Flexible tubules, Gradient fractions, Gumboro disease, Hexagonal arrangement, Hexagonal lattices, Ibdv, Ibdv capsids, Ibdv orfa1, Ibdv particles, Ibdv polyprotein, Ibdv virions, Ibdv vlps, Icosahedral, Icosahedral caps, Icosahedral symmetry, Immunoblotting analysis, Immunoelectron microscopy, Infectious bursal disease, Infectious bursal disease virus, Inner surfaces, Insect cells, Jackson immunoresearch laboratories, Lattice, Long tubules, Longitudinal sections, Molecular mass marker sizes, Monospecific, Monospecific antisera, Morphogenesis, Negative staining, Neutralizing antibodies, Nitrocellulose membranes, Other fractions, Pacym1, Polyclonal, Polyclonal rabbit, Polypeptide, Polyprotein, Precursor polyprotein, Previous reports, Recombinant, Recombinant baculovirus, Recombinant baculoviruses, Rigid tubules, Rigid type, Rodriguez, Room temperature, Spherical vlps, Standard methods, Structural analysis, Structural characterization, Structural proteins, Sucrose, Sucrose cushion, Sucrose gradient, Transmissible gastroenteritis coronavirus, Trimer, Tubular structures, Tubule, Type tubules, Ultrathin sections, Uranyl acetate, Viral capsid, Virion, Virol, Vlps, Western blot analysis.
Abstract
Abstract: Infectious bursal disease virus (IBDV) capsid is formed by the processing of a large polyprotein and subsequent assembly of VPX/VP2 and VP3. To learn more about the processing of the polyprotein and factors affecting the correct assembly of the viral capsid in vitro, different constructs were made using two baculovirus transfer vectors, pFastBac and pAcYM1. Surprisingly, the expression of the capsid proteins gave rise to different types of particles in each system, as observed by electron microscopy and immunofluorescence. FastBac expression led to the production of only rigid tubular structures, similar to those described as type I in viral infection. Western blot analysis revealed that these rigid tubules are formed exclusively by VPX. These tubules revealed a hexagonal arrangement of units that are trimer clustered, similar to those observed in IBDV virions. In contrast, pAcYM1 expression led to the assembly of virus-like particles (VLPs), flexible tubules, and intermediate assembly products formed by icosahedral caps elongated in tubes, suggesting an aberrant morphogenesis. Processing of VPX to VP2 seems to be a crucial requirement for the proper morphogenesis and assembly of IBDV particles. After immunoelectron microscopy, VPX/VP2 was detected on the surface of tubules and VLPs. We also demonstrated that VP3 is found only on the inner surfaces of VLPs and caps of the tubular structures. In summary, assembly of VLPs requires the internal scaffolding of VP3, which seems to induce the closing of the tubular architecture into VLPs and, thereafter, the subsequent processing of VPX to VP2.
Url:
DOI: 10.1006/viro.2000.0559
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 001D81
- to stream Istex, to step Curation: 001D81
- to stream Istex, to step Checkpoint: 001047
- to stream Main, to step Merge: 003790
- to stream Main, to step Curation: 003748
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Different Architectures in the Assembly of Infectious Bursal Disease Virus Capsid Proteins Expressed in Insect Cells</title><author><name sortKey="Martinez Torrecuadrada, Jorge Luis" sort="Martinez Torrecuadrada, Jorge Luis" uniqKey="Martinez Torrecuadrada J" first="Jorge Luis" last="Martinez-Torrecuadrada">Jorge Luis Martinez-Torrecuadrada</name></author><author><name sortKey="Cast N, Jose R" sort="Cast N, Jose R" uniqKey="Cast N J" first="José R." last="Cast N">José R. Cast N</name></author><author><name sortKey="Castro, Marian" sort="Castro, Marian" uniqKey="Castro M" first="Marian" last="Castro">Marian Castro</name></author><author><name sortKey="Carrascosa, Jose L" sort="Carrascosa, Jose L" uniqKey="Carrascosa J" first="José L." last="Carrascosa">José L. Carrascosa</name></author><author><name sortKey="Rodriguez, Jose F" sort="Rodriguez, Jose F" uniqKey="Rodriguez J" first="José F." last="Rodriguez">José F. Rodriguez</name></author><author><name sortKey="Casal, J Ignacio" sort="Casal, J Ignacio" uniqKey="Casal J" first="J. Ignacio" last="Casal">J. Ignacio Casal</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:595D241D72657B86E4E07878A40827C71CC5A801</idno><date when="2000" year="2000">2000</date><idno type="doi">10.1006/viro.2000.0559</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-Q0V5WTFX-Z/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">001D81</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">001D81</idno><idno type="wicri:Area/Istex/Curation">001D81</idno><idno type="wicri:Area/Istex/Checkpoint">001047</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001047</idno><idno type="wicri:doubleKey">0042-6822:2000:Martinez Torrecuadrada J:different:architectures:in</idno><idno type="wicri:Area/Main/Merge">003790</idno><idno type="wicri:Area/Main/Curation">003748</idno><idno type="wicri:Area/Main/Exploration">003748</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Different Architectures in the Assembly of Infectious Bursal Disease Virus Capsid Proteins Expressed in Insect Cells</title><author><name sortKey="Martinez Torrecuadrada, Jorge Luis" sort="Martinez Torrecuadrada, Jorge Luis" uniqKey="Martinez Torrecuadrada J" first="Jorge Luis" last="Martinez-Torrecuadrada">Jorge Luis Martinez-Torrecuadrada</name><affiliation wicri:level="3"><country xml:lang="fr">Espagne</country><wicri:regionArea>INGENASA, C/Hnos, Garcia Noblejas 41, 4° 28037, Madrid</wicri:regionArea><placeName><settlement type="city">Madrid</settlement><region nuts="2" type="region">Communauté de Madrid</region></placeName></affiliation></author><author><name sortKey="Cast N, Jose R" sort="Cast N, Jose R" uniqKey="Cast N J" first="José R." last="Cast N">José R. Cast N</name><affiliation wicri:level="3"><country xml:lang="fr">Espagne</country><wicri:regionArea>Departamento de Estructura de Macromoléculas, Centro Nacional de Biotecnologı́a, Cantoblanco, 28049, Madrid</wicri:regionArea><placeName><settlement type="city">Madrid</settlement><region nuts="2" type="region">Communauté de Madrid</region></placeName></affiliation></author><author><name sortKey="Castro, Marian" sort="Castro, Marian" uniqKey="Castro M" first="Marian" last="Castro">Marian Castro</name><affiliation wicri:level="3"><country xml:lang="fr">Espagne</country><wicri:regionArea>Departamento de Biologı́a Molecular y Celular, Centro Nacional de Biotecnologı́a, Cantoblanco, 28049, Madrid</wicri:regionArea><placeName><settlement type="city">Madrid</settlement><region nuts="2" type="region">Communauté de Madrid</region></placeName></affiliation></author><author><name sortKey="Carrascosa, Jose L" sort="Carrascosa, Jose L" uniqKey="Carrascosa J" first="José L." last="Carrascosa">José L. Carrascosa</name><affiliation wicri:level="3"><country xml:lang="fr">Espagne</country><wicri:regionArea>Departamento de Estructura de Macromoléculas, Centro Nacional de Biotecnologı́a, Cantoblanco, 28049, Madrid</wicri:regionArea><placeName><settlement type="city">Madrid</settlement><region nuts="2" type="region">Communauté de Madrid</region></placeName></affiliation></author><author><name sortKey="Rodriguez, Jose F" sort="Rodriguez, Jose F" uniqKey="Rodriguez J" first="José F." last="Rodriguez">José F. Rodriguez</name><affiliation wicri:level="3"><country xml:lang="fr">Espagne</country><wicri:regionArea>Departamento de Biologı́a Molecular y Celular, Centro Nacional de Biotecnologı́a, Cantoblanco, 28049, Madrid</wicri:regionArea><placeName><settlement type="city">Madrid</settlement><region nuts="2" type="region">Communauté de Madrid</region></placeName></affiliation></author><author><name sortKey="Casal, J Ignacio" sort="Casal, J Ignacio" uniqKey="Casal J" first="J. Ignacio" last="Casal">J. Ignacio Casal</name><affiliation wicri:level="3"><country xml:lang="fr">Espagne</country><wicri:regionArea>INGENASA, C/Hnos, Garcia Noblejas 41, 4° 28037, Madrid</wicri:regionArea><placeName><settlement type="city">Madrid</settlement><region nuts="2" type="region">Communauté de Madrid</region></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">Virology</title><title level="j" type="abbrev">YVIRO</title><idno type="ISSN">0042-6822</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="2000">2000</date><biblScope unit="volume">278</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="322">322</biblScope><biblScope unit="page" to="331">331</biblScope></imprint><idno type="ISSN">0042-6822</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0042-6822</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Antigenic properties</term><term>Antiserum</term><term>Assembly products</term><term>Baculovirus</term><term>Baculovirus transfer vectors</term><term>Baculovirus vectors</term><term>Bluetongue virus</term><term>Bursal</term><term>Capsid</term><term>Capsid protein</term><term>Capsomere lattice</term><term>Carrascosa</term><term>Cellular extracts</term><term>Confocal</term><term>Confocal microscopy</term><term>Different architectures</term><term>Different baculovirus transfer vectors</term><term>Different types</term><term>Electron microscopy</term><term>Electron microscopy analysis</term><term>Escherichia coli</term><term>Flexible tubules</term><term>Gradient fractions</term><term>Gumboro disease</term><term>Hexagonal arrangement</term><term>Hexagonal lattices</term><term>Ibdv</term><term>Ibdv capsids</term><term>Ibdv orfa1</term><term>Ibdv particles</term><term>Ibdv polyprotein</term><term>Ibdv virions</term><term>Ibdv vlps</term><term>Icosahedral</term><term>Icosahedral caps</term><term>Icosahedral symmetry</term><term>Immunoblotting analysis</term><term>Immunoelectron microscopy</term><term>Infectious bursal disease</term><term>Infectious bursal disease virus</term><term>Inner surfaces</term><term>Insect cells</term><term>Jackson immunoresearch laboratories</term><term>Lattice</term><term>Long tubules</term><term>Longitudinal sections</term><term>Molecular mass marker sizes</term><term>Monospecific</term><term>Monospecific antisera</term><term>Morphogenesis</term><term>Negative staining</term><term>Neutralizing antibodies</term><term>Nitrocellulose membranes</term><term>Other fractions</term><term>Pacym1</term><term>Polyclonal</term><term>Polyclonal rabbit</term><term>Polypeptide</term><term>Polyprotein</term><term>Precursor polyprotein</term><term>Previous reports</term><term>Recombinant</term><term>Recombinant baculovirus</term><term>Recombinant baculoviruses</term><term>Rigid tubules</term><term>Rigid type</term><term>Rodriguez</term><term>Room temperature</term><term>Spherical vlps</term><term>Standard methods</term><term>Structural analysis</term><term>Structural characterization</term><term>Structural proteins</term><term>Sucrose</term><term>Sucrose cushion</term><term>Sucrose gradient</term><term>Transmissible gastroenteritis coronavirus</term><term>Trimer</term><term>Tubular structures</term><term>Tubule</term><term>Type tubules</term><term>Ultrathin sections</term><term>Uranyl acetate</term><term>Viral capsid</term><term>Virion</term><term>Virol</term><term>Vlps</term><term>Western blot analysis</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Infectious bursal disease virus (IBDV) capsid is formed by the processing of a large polyprotein and subsequent assembly of VPX/VP2 and VP3. To learn more about the processing of the polyprotein and factors affecting the correct assembly of the viral capsid in vitro, different constructs were made using two baculovirus transfer vectors, pFastBac and pAcYM1. Surprisingly, the expression of the capsid proteins gave rise to different types of particles in each system, as observed by electron microscopy and immunofluorescence. FastBac expression led to the production of only rigid tubular structures, similar to those described as type I in viral infection. Western blot analysis revealed that these rigid tubules are formed exclusively by VPX. These tubules revealed a hexagonal arrangement of units that are trimer clustered, similar to those observed in IBDV virions. In contrast, pAcYM1 expression led to the assembly of virus-like particles (VLPs), flexible tubules, and intermediate assembly products formed by icosahedral caps elongated in tubes, suggesting an aberrant morphogenesis. Processing of VPX to VP2 seems to be a crucial requirement for the proper morphogenesis and assembly of IBDV particles. After immunoelectron microscopy, VPX/VP2 was detected on the surface of tubules and VLPs. We also demonstrated that VP3 is found only on the inner surfaces of VLPs and caps of the tubular structures. In summary, assembly of VLPs requires the internal scaffolding of VP3, which seems to induce the closing of the tubular architecture into VLPs and, thereafter, the subsequent processing of VPX to VP2.</div></front></TEI><affiliations><list><country><li>Espagne</li></country><region><li>Communauté de Madrid</li></region><settlement><li>Madrid</li></settlement></list><tree><country name="Espagne"><region name="Communauté de Madrid"><name sortKey="Martinez Torrecuadrada, Jorge Luis" sort="Martinez Torrecuadrada, Jorge Luis" uniqKey="Martinez Torrecuadrada J" first="Jorge Luis" last="Martinez-Torrecuadrada">Jorge Luis Martinez-Torrecuadrada</name></region><name sortKey="Carrascosa, Jose L" sort="Carrascosa, Jose L" uniqKey="Carrascosa J" first="José L." last="Carrascosa">José L. Carrascosa</name><name sortKey="Casal, J Ignacio" sort="Casal, J Ignacio" uniqKey="Casal J" first="J. Ignacio" last="Casal">J. Ignacio Casal</name><name sortKey="Cast N, Jose R" sort="Cast N, Jose R" uniqKey="Cast N J" first="José R." last="Cast N">José R. Cast N</name><name sortKey="Castro, Marian" sort="Castro, Marian" uniqKey="Castro M" first="Marian" last="Castro">Marian Castro</name><name sortKey="Rodriguez, Jose F" sort="Rodriguez, Jose F" uniqKey="Rodriguez J" first="José F." last="Rodriguez">José F. Rodriguez</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 003748 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 003748 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= ISTEX:595D241D72657B86E4E07878A40827C71CC5A801
|texte= Different Architectures in the Assembly of Infectious Bursal Disease Virus Capsid Proteins Expressed in Insect Cells
}}
| This area was generated with Dilib version V0.6.33. |  |