Role of endocytosis and low pH in murine hepatitis virus strain A59 cell entry.
Identifieur interne : 003611 ( Main/Exploration ); précédent : 003610; suivant : 003612Role of endocytosis and low pH in murine hepatitis virus strain A59 cell entry.
Auteurs : Patricia Eifart [Allemagne] ; Kai Ludwig ; Christoph Böttcher ; Cornelis A M. De Haan ; Peter J M. Rottier ; Thomas Korte ; Andreas HerrmannSource :
- Journal of virology [ 0022-538X ] ; 2007.
Descripteurs français
- KwdFr :
- Animaux, Concentration en ions d'hydrogène, Conformation des protéines, Endocytose (), Fibroblastes (virologie), Fusion membranaire, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (), Glycoprotéines membranaires (métabolisme), Inhibiteurs de protéases (pharmacologie), Lignée cellulaire, Lysosomes (), Lysosomes (virologie), Protéines de l'enveloppe virale (), Protéines de l'enveloppe virale (métabolisme), Pénétration virale (), Souris, Spectrométrie de fluorescence, Virus de l'hépatite murine (physiologie).
- MESH :
- métabolisme : Glycoprotéines membranaires, Protéines de l'enveloppe virale.
- pharmacologie : Inhibiteurs de protéases.
- physiologie : Virus de l'hépatite murine.
- virologie : Fibroblastes, Lysosomes.
- Animaux, Concentration en ions d'hydrogène, Conformation des protéines, Endocytose, Fusion membranaire, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires, Lignée cellulaire, Lysosomes, Protéines de l'enveloppe virale, Pénétration virale, Souris, Spectrométrie de fluorescence.
English descriptors
- KwdEn :
- Animals, Cell Line, Endocytosis (drug effects), Fibroblasts (virology), Hydrogen-Ion Concentration, Lysosomes (drug effects), Lysosomes (virology), Membrane Fusion, Membrane Glycoproteins (chemistry), Membrane Glycoproteins (metabolism), Mice, Murine hepatitis virus (physiology), Protease Inhibitors (pharmacology), Protein Conformation, Spectrometry, Fluorescence, Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (chemistry), Viral Envelope Proteins (metabolism), Virus Internalization (drug effects).
- MESH :
- chemical , chemistry : Membrane Glycoproteins, Viral Envelope Proteins.
- drug effects : Endocytosis, Lysosomes, Virus Internalization.
- chemical , metabolism : Membrane Glycoproteins, Viral Envelope Proteins.
- chemical , pharmacology : Protease Inhibitors.
- physiology : Murine hepatitis virus.
- virology : Fibroblasts, Lysosomes.
- Animals, Cell Line, Hydrogen-Ion Concentration, Membrane Fusion, Mice, Protein Conformation, Spectrometry, Fluorescence, Spike Glycoprotein, Coronavirus.
Abstract
Infection by the coronavirus mouse hepatitis virus strain A59 (MHV-A59) requires the release of the viral genome by fusion with the respective target membrane of the host cell. Fusion is mediated by the viral S protein. Here, the entry pathway of MHV-A59 into murine fibroblast cells was studied by independent approaches. Infection of cells assessed by plaque reduction assay was strongly inhibited by lysosomotropic compounds and substances that interfere with clathrin-dependent endocytosis, suggesting that MHV-A59 is taken up via endocytosis and delivered to acidic endosomal compartments. Infection was only slightly reduced in the presence of substances inhibiting proteases of endosomal compartments, precluding that the endocytic uptake is required to activate the fusion potential of the S protein by its cleavage. Fluorescence confocal microscopy of labeled MHV-A59 confirmed that virus is taken up via endocytosis. Bright labeling of intracellular compartments suggests their fusion with the viral envelope. No fusion with the plasma membrane was observed at neutral pH conditions. However, when virus was bound to cells and the pH was lowered to 5.0, we observed a strong labeling of the plasma membrane. Electron microscopy revealed low pH triggered conformational alterations of the S ectodomain. Very likely, these alterations are irreversible because low-pH treatment of viruses in the absence of target membranes caused an irreversible loss of the fusion activity. The results imply that endocytosis plays a major role in MHV-A59 infection and the acidic pH of the endosomal compartment triggers a conformational change of the S protein mediating fusion.
DOI: 10.1128/JVI.00725-07
PubMed: 17626088
Affiliations:
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Le document en format XML
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Cell Line</term>
<term>Endocytosis (drug effects)</term>
<term>Fibroblasts (virology)</term>
<term>Hydrogen-Ion Concentration</term>
<term>Lysosomes (drug effects)</term>
<term>Lysosomes (virology)</term>
<term>Membrane Fusion</term>
<term>Membrane Glycoproteins (chemistry)</term>
<term>Membrane Glycoproteins (metabolism)</term>
<term>Mice</term>
<term>Murine hepatitis virus (physiology)</term>
<term>Protease Inhibitors (pharmacology)</term>
<term>Protein Conformation</term>
<term>Spectrometry, Fluorescence</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Viral Envelope Proteins (chemistry)</term>
<term>Viral Envelope Proteins (metabolism)</term>
<term>Virus Internalization (drug effects)</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term>
<term>Concentration en ions d'hydrogène</term>
<term>Conformation des protéines</term>
<term>Endocytose ()</term>
<term>Fibroblastes (virologie)</term>
<term>Fusion membranaire</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires ()</term>
<term>Glycoprotéines membranaires (métabolisme)</term>
<term>Inhibiteurs de protéases (pharmacologie)</term>
<term>Lignée cellulaire</term>
<term>Lysosomes ()</term>
<term>Lysosomes (virologie)</term>
<term>Protéines de l'enveloppe virale ()</term>
<term>Protéines de l'enveloppe virale (métabolisme)</term>
<term>Pénétration virale ()</term>
<term>Souris</term>
<term>Spectrométrie de fluorescence</term>
<term>Virus de l'hépatite murine (physiologie)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Membrane Glycoproteins</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Endocytosis</term>
<term>Lysosomes</term>
<term>Virus Internalization</term>
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<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Membrane Glycoproteins</term>
<term>Viral Envelope Proteins</term>
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<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
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<keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Inhibiteurs de protéases</term>
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<keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Protease Inhibitors</term>
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<keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Virus de l'hépatite murine</term>
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<keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Murine hepatitis virus</term>
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<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Fibroblastes</term>
<term>Lysosomes</term>
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<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Fibroblasts</term>
<term>Lysosomes</term>
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<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Cell Line</term>
<term>Hydrogen-Ion Concentration</term>
<term>Membrane Fusion</term>
<term>Mice</term>
<term>Protein Conformation</term>
<term>Spectrometry, Fluorescence</term>
<term>Spike Glycoprotein, Coronavirus</term>
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<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Concentration en ions d'hydrogène</term>
<term>Conformation des protéines</term>
<term>Endocytose</term>
<term>Fusion membranaire</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires</term>
<term>Lignée cellulaire</term>
<term>Lysosomes</term>
<term>Protéines de l'enveloppe virale</term>
<term>Pénétration virale</term>
<term>Souris</term>
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<front><div type="abstract" xml:lang="en">Infection by the coronavirus mouse hepatitis virus strain A59 (MHV-A59) requires the release of the viral genome by fusion with the respective target membrane of the host cell. Fusion is mediated by the viral S protein. Here, the entry pathway of MHV-A59 into murine fibroblast cells was studied by independent approaches. Infection of cells assessed by plaque reduction assay was strongly inhibited by lysosomotropic compounds and substances that interfere with clathrin-dependent endocytosis, suggesting that MHV-A59 is taken up via endocytosis and delivered to acidic endosomal compartments. Infection was only slightly reduced in the presence of substances inhibiting proteases of endosomal compartments, precluding that the endocytic uptake is required to activate the fusion potential of the S protein by its cleavage. Fluorescence confocal microscopy of labeled MHV-A59 confirmed that virus is taken up via endocytosis. Bright labeling of intracellular compartments suggests their fusion with the viral envelope. No fusion with the plasma membrane was observed at neutral pH conditions. However, when virus was bound to cells and the pH was lowered to 5.0, we observed a strong labeling of the plasma membrane. Electron microscopy revealed low pH triggered conformational alterations of the S ectodomain. Very likely, these alterations are irreversible because low-pH treatment of viruses in the absence of target membranes caused an irreversible loss of the fusion activity. The results imply that endocytosis plays a major role in MHV-A59 infection and the acidic pH of the endosomal compartment triggers a conformational change of the S protein mediating fusion.</div>
</front>
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<tree><noCountry><name sortKey="Bottcher, Christoph" sort="Bottcher, Christoph" uniqKey="Bottcher C" first="Christoph" last="Böttcher">Christoph Böttcher</name>
<name sortKey="De Haan, Cornelis A M" sort="De Haan, Cornelis A M" uniqKey="De Haan C" first="Cornelis A M" last="De Haan">Cornelis A M. De Haan</name>
<name sortKey="Herrmann, Andreas" sort="Herrmann, Andreas" uniqKey="Herrmann A" first="Andreas" last="Herrmann">Andreas Herrmann</name>
<name sortKey="Korte, Thomas" sort="Korte, Thomas" uniqKey="Korte T" first="Thomas" last="Korte">Thomas Korte</name>
<name sortKey="Ludwig, Kai" sort="Ludwig, Kai" uniqKey="Ludwig K" first="Kai" last="Ludwig">Kai Ludwig</name>
<name sortKey="Rottier, Peter J M" sort="Rottier, Peter J M" uniqKey="Rottier P" first="Peter J M" last="Rottier">Peter J M. Rottier</name>
</noCountry>
<country name="Allemagne"><region name="Berlin"><name sortKey="Eifart, Patricia" sort="Eifart, Patricia" uniqKey="Eifart P" first="Patricia" last="Eifart">Patricia Eifart</name>
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