Characterization of a Highly Conserved Domain within the Severe Acute Respiratory Syndrome Coronavirus Spike Protein S2 Domain with Characteristics of a Viral Fusion Peptide▿
Identifieur interne : 001F17 ( Ncbi/Merge ); précédent : 001F16; suivant : 001F18Characterization of a Highly Conserved Domain within the Severe Acute Respiratory Syndrome Coronavirus Spike Protein S2 Domain with Characteristics of a Viral Fusion Peptide▿
Auteurs : Ikenna G. Madu [États-Unis] ; Shoshannah L. Roth [États-Unis] ; Sandrine Belouzard [États-Unis] ; Gary R. Whittaker [États-Unis]Source :
- Journal of Virology [ 0022-538X ] ; 2009.
Descripteurs français
- KwdFr :
- Alignement de séquences, Animaux, Données de séquences moléculaires, Fusion cellulaire, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (), Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (physiologie), Humains, Lignée cellulaire, Peptides (), Peptides (génétique), Peptides (métabolisme), Protéines de l'enveloppe virale (), Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (physiologie), Pénétration virale, Structure tertiaire des protéines, Syndrome respiratoire aigu sévère (virologie), Séquence d'acides aminés, Virus du SRAS (), Virus du SRAS (génétique), Virus du SRAS (métabolisme).
- MESH :
- génétique : Glycoprotéines membranaires, Peptides, Protéines de l'enveloppe virale, Virus du SRAS.
- métabolisme : Peptides, Virus du SRAS.
- physiologie : Glycoprotéines membranaires, Protéines de l'enveloppe virale.
- virologie : Syndrome respiratoire aigu sévère.
- Alignement de séquences, Animaux, Données de séquences moléculaires, Fusion cellulaire, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires, Humains, Lignée cellulaire, Peptides, Protéines de l'enveloppe virale, Pénétration virale, Structure tertiaire des protéines, Séquence d'acides aminés, Virus du SRAS.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, Cell Fusion, Cell Line, Humans, Membrane Glycoproteins (chemistry), Membrane Glycoproteins (genetics), Membrane Glycoproteins (physiology), Molecular Sequence Data, Peptides (chemistry), Peptides (genetics), Peptides (metabolism), Protein Structure, Tertiary, SARS Virus (chemistry), SARS Virus (genetics), SARS Virus (metabolism), Sequence Alignment, Severe Acute Respiratory Syndrome (virology), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (chemistry), Viral Envelope Proteins (genetics), Viral Envelope Proteins (physiology), Virus Internalization.
- MESH :
- chemical , chemistry : Membrane Glycoproteins, Peptides, Viral Envelope Proteins.
- chemical , genetics : Membrane Glycoproteins, Peptides, Viral Envelope Proteins.
- chemical , metabolism : Peptides.
- chemical , physiology : Membrane Glycoproteins, Viral Envelope Proteins.
- chemistry : SARS Virus.
- genetics : SARS Virus.
- metabolism : SARS Virus.
- virology : Severe Acute Respiratory Syndrome.
- Amino Acid Sequence, Animals, Cell Fusion, Cell Line, Humans, Molecular Sequence Data, Protein Structure, Tertiary, Sequence Alignment, Spike Glycoprotein, Coronavirus, Virus Internalization.
Abstract
Many viral fusion proteins are primed by proteolytic cleavage near their fusion peptides. While the coronavirus (CoV) spike (S) protein is known to be cleaved at the S1/S2 boundary, this cleavage site is not closely linked to a fusion peptide. However, a second cleavage site has been identified in the severe acute respiratory syndrome CoV (SARS-CoV) S2 domain (R797). Here, we investigated whether this internal cleavage of S2 exposes a viral fusion peptide. We show that the residues immediately C-terminal to the SARS-CoV S2 cleavage site SFIEDLLFNKVTLADAGF are very highly conserved across all CoVs. Mutagenesis studies of these residues in SARS-CoV S, followed by cell-cell fusion and pseudotyped virion infectivity assays, showed a critical role for residues L803, L804, and F805 in membrane fusion. Mutation of the most N-terminal residue (S798) had little or no effect on membrane fusion. Biochemical analyses of synthetic peptides corresponding to the proposed S2 fusion peptide also showed an important role for this region in membrane fusion and indicated the presence of α-helical structure. We propose that proteolytic cleavage within S2 exposes a novel internal fusion peptide for SARS-CoV S, which may be conserved across the
Url:
DOI: 10.1128/JVI.00079-09
PubMed: 19439480
PubMed Central: 2708636
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PMC:2708636Le document en format XML
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(physiology)</term><term>Molecular Sequence Data</term><term>Peptides (chemistry)</term><term>Peptides (genetics)</term><term>Peptides (metabolism)</term><term>Protein Structure, Tertiary</term><term>SARS Virus (chemistry)</term><term>SARS Virus (genetics)</term><term>SARS Virus (metabolism)</term><term>Sequence Alignment</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (chemistry)</term><term>Viral Envelope Proteins (genetics)</term><term>Viral Envelope Proteins (physiology)</term><term>Virus Internalization</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Alignement de séquences</term><term>Animaux</term><term>Données de séquences moléculaires</term><term>Fusion cellulaire</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires ()</term><term>Glycoprotéines membranaires (génétique)</term><term>Glycoprotéines membranaires 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type="chemical" qualifier="metabolism" xml:lang="en"><term>Peptides</term></keywords><keywords scheme="MESH" type="chemical" qualifier="physiology" xml:lang="en"><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Peptides</term><term>Protéines de l'enveloppe virale</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Peptides</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Cell Fusion</term><term>Cell Line</term><term>Humans</term><term>Molecular Sequence Data</term><term>Protein Structure, Tertiary</term><term>Sequence Alignment</term><term>Spike Glycoprotein, Coronavirus</term><term>Virus Internalization</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Alignement de séquences</term><term>Animaux</term><term>Données de séquences moléculaires</term><term>Fusion cellulaire</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires</term><term>Humains</term><term>Lignée cellulaire</term><term>Peptides</term><term>Protéines de l'enveloppe virale</term><term>Pénétration virale</term><term>Structure tertiaire des protéines</term><term>Séquence d'acides aminés</term><term>Virus du SRAS</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Many viral fusion proteins are primed by proteolytic cleavage near their fusion peptides. While the coronavirus (CoV) spike (S) protein is known to be cleaved at the S1/S2 boundary, this cleavage site is not closely linked to a fusion peptide. However, a second cleavage site has been identified in the severe acute respiratory syndrome CoV (SARS-CoV) S2 domain (R797). Here, we investigated whether this internal cleavage of S2 exposes a viral fusion peptide. We show that the residues immediately C-terminal to the SARS-CoV S2 cleavage site SFIEDLLFNKVTLADAGF are very highly conserved across all CoVs. Mutagenesis studies of these residues in SARS-CoV S, followed by cell-cell fusion and pseudotyped virion infectivity assays, showed a critical role for residues L803, L804, and F805 in membrane fusion. Mutation of the most N-terminal residue (S798) had little or no effect on membrane fusion. Biochemical analyses of synthetic peptides corresponding to the proposed S2 fusion peptide also showed an important role for this region in membrane fusion and indicated the presence of α-helical structure. We propose that proteolytic cleavage within S2 exposes a novel internal fusion peptide for SARS-CoV S, which may be conserved across the <italic>Coronaviridae</italic>.</p></div></front></TEI><double pmid="19439480"><pmc><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Characterization of a Highly Conserved Domain within the Severe Acute Respiratory Syndrome Coronavirus Spike Protein S2 Domain with Characteristics of a Viral Fusion Peptide<xref ref-type="fn" rid="fn1">▿</xref> </title><author><name sortKey="Madu, Ikenna G" sort="Madu, Ikenna G" uniqKey="Madu I" first="Ikenna G." last="Madu">Ikenna G. Madu</name><affiliation wicri:level="2"><nlm:aff id="aff0">Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca</wicri:cityArea></affiliation></author><author><name sortKey="Roth, Shoshannah L" sort="Roth, Shoshannah L" uniqKey="Roth S" first="Shoshannah L." last="Roth">Shoshannah L. Roth</name><affiliation wicri:level="2"><nlm:aff id="aff0">Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca</wicri:cityArea></affiliation></author><author><name sortKey="Belouzard, Sandrine" sort="Belouzard, Sandrine" uniqKey="Belouzard S" first="Sandrine" last="Belouzard">Sandrine Belouzard</name><affiliation wicri:level="2"><nlm:aff id="aff0">Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca</wicri:cityArea></affiliation></author><author><name sortKey="Whittaker, Gary R" sort="Whittaker, Gary R" uniqKey="Whittaker G" first="Gary R." last="Whittaker">Gary R. Whittaker</name><affiliation wicri:level="2"><nlm:aff id="aff0">Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca</wicri:cityArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">19439480</idno><idno type="pmc">2708636</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2708636</idno><idno type="RBID">PMC:2708636</idno><idno type="doi">10.1128/JVI.00079-09</idno><date when="2009">2009</date><idno type="wicri:Area/Pmc/Corpus">000674</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000674</idno><idno type="wicri:Area/Pmc/Curation">000674</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000674</idno><idno type="wicri:Area/Pmc/Checkpoint">000D41</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000D41</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Characterization of a Highly Conserved Domain within the Severe Acute Respiratory Syndrome Coronavirus Spike Protein S2 Domain with Characteristics of a Viral Fusion Peptide<xref ref-type="fn" rid="fn1">▿</xref> </title><author><name sortKey="Madu, Ikenna G" sort="Madu, Ikenna G" uniqKey="Madu I" first="Ikenna G." last="Madu">Ikenna G. Madu</name><affiliation wicri:level="2"><nlm:aff id="aff0">Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca</wicri:cityArea></affiliation></author><author><name sortKey="Roth, Shoshannah L" sort="Roth, Shoshannah L" uniqKey="Roth S" first="Shoshannah L." last="Roth">Shoshannah L. Roth</name><affiliation wicri:level="2"><nlm:aff id="aff0">Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca</wicri:cityArea></affiliation></author><author><name sortKey="Belouzard, Sandrine" sort="Belouzard, Sandrine" uniqKey="Belouzard S" first="Sandrine" last="Belouzard">Sandrine Belouzard</name><affiliation wicri:level="2"><nlm:aff id="aff0">Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca</wicri:cityArea></affiliation></author><author><name sortKey="Whittaker, Gary R" sort="Whittaker, Gary R" uniqKey="Whittaker G" first="Gary R." last="Whittaker">Gary R. Whittaker</name><affiliation wicri:level="2"><nlm:aff id="aff0">Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">État de New York</region></placeName><wicri:cityArea>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca</wicri:cityArea></affiliation></author></analytic><series><title level="j">Journal of Virology</title><idno type="ISSN">0022-538X</idno><idno type="eISSN">1098-5514</idno><imprint><date when="2009">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Many viral fusion proteins are primed by proteolytic cleavage near their fusion peptides. While the coronavirus (CoV) spike (S) protein is known to be cleaved at the S1/S2 boundary, this cleavage site is not closely linked to a fusion peptide. However, a second cleavage site has been identified in the severe acute respiratory syndrome CoV (SARS-CoV) S2 domain (R797). Here, we investigated whether this internal cleavage of S2 exposes a viral fusion peptide. We show that the residues immediately C-terminal to the SARS-CoV S2 cleavage site SFIEDLLFNKVTLADAGF are very highly conserved across all CoVs. Mutagenesis studies of these residues in SARS-CoV S, followed by cell-cell fusion and pseudotyped virion infectivity assays, showed a critical role for residues L803, L804, and F805 in membrane fusion. Mutation of the most N-terminal residue (S798) had little or no effect on membrane fusion. Biochemical analyses of synthetic peptides corresponding to the proposed S2 fusion peptide also showed an important role for this region in membrane fusion and indicated the presence of α-helical structure. We propose that proteolytic cleavage within S2 exposes a novel internal fusion peptide for SARS-CoV S, which may be conserved across the <italic>Coronaviridae</italic>.</p></div></front></TEI></pmc><pubmed><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Characterization of a highly conserved domain within the severe acute respiratory syndrome coronavirus spike protein S2 domain with characteristics of a viral fusion peptide.</title><author><name sortKey="Madu, Ikenna G" sort="Madu, Ikenna G" uniqKey="Madu I" first="Ikenna G" last="Madu">Ikenna G. Madu</name><affiliation wicri:level="4"><nlm:affiliation>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853</wicri:regionArea><placeName><region type="state">État de New York</region><settlement type="city">Ithaca (New York)</settlement></placeName><orgName type="university">Université Cornell</orgName></affiliation></author><author><name sortKey="Roth, Shoshannah L" sort="Roth, Shoshannah L" uniqKey="Roth S" first="Shoshannah L" last="Roth">Shoshannah L. Roth</name></author><author><name sortKey="Belouzard, Sandrine" sort="Belouzard, Sandrine" uniqKey="Belouzard S" first="Sandrine" last="Belouzard">Sandrine Belouzard</name></author><author><name sortKey="Whittaker, Gary R" sort="Whittaker, Gary R" uniqKey="Whittaker G" first="Gary R" last="Whittaker">Gary R. Whittaker</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2009">2009</date><idno type="RBID">pubmed:19439480</idno><idno type="pmid">19439480</idno><idno type="doi">10.1128/JVI.00079-09</idno><idno type="wicri:Area/PubMed/Corpus">001900</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001900</idno><idno type="wicri:Area/PubMed/Curation">001900</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001900</idno><idno type="wicri:Area/PubMed/Checkpoint">001929</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001929</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Characterization of a highly conserved domain within the severe acute respiratory syndrome coronavirus spike protein S2 domain with characteristics of a viral fusion peptide.</title><author><name sortKey="Madu, Ikenna G" sort="Madu, Ikenna G" uniqKey="Madu I" first="Ikenna G" last="Madu">Ikenna G. Madu</name><affiliation wicri:level="4"><nlm:affiliation>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853</wicri:regionArea><placeName><region type="state">État de New York</region><settlement type="city">Ithaca (New York)</settlement></placeName><orgName type="university">Université Cornell</orgName></affiliation></author><author><name sortKey="Roth, Shoshannah L" sort="Roth, Shoshannah L" uniqKey="Roth S" first="Shoshannah L" last="Roth">Shoshannah L. Roth</name></author><author><name sortKey="Belouzard, Sandrine" sort="Belouzard, Sandrine" uniqKey="Belouzard S" first="Sandrine" last="Belouzard">Sandrine Belouzard</name></author><author><name sortKey="Whittaker, Gary R" sort="Whittaker, Gary R" uniqKey="Whittaker G" first="Gary R" last="Whittaker">Gary R. Whittaker</name></author></analytic><series><title level="j">Journal of virology</title><idno type="eISSN">1098-5514</idno><imprint><date when="2009" type="published">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Cell Fusion</term><term>Cell Line</term><term>Humans</term><term>Membrane Glycoproteins (chemistry)</term><term>Membrane Glycoproteins (genetics)</term><term>Membrane Glycoproteins (physiology)</term><term>Molecular Sequence Data</term><term>Peptides (chemistry)</term><term>Peptides (genetics)</term><term>Peptides (metabolism)</term><term>Protein Structure, Tertiary</term><term>SARS Virus (chemistry)</term><term>SARS Virus (genetics)</term><term>SARS Virus (metabolism)</term><term>Sequence Alignment</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (chemistry)</term><term>Viral Envelope Proteins (genetics)</term><term>Viral Envelope Proteins (physiology)</term><term>Virus Internalization</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Alignement de séquences</term><term>Animaux</term><term>Données de séquences moléculaires</term><term>Fusion cellulaire</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires ()</term><term>Glycoprotéines membranaires (génétique)</term><term>Glycoprotéines membranaires (physiologie)</term><term>Humains</term><term>Lignée cellulaire</term><term>Peptides ()</term><term>Peptides (génétique)</term><term>Peptides (métabolisme)</term><term>Protéines de l'enveloppe virale ()</term><term>Protéines de l'enveloppe virale (génétique)</term><term>Protéines de l'enveloppe virale (physiologie)</term><term>Pénétration virale</term><term>Structure tertiaire des protéines</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Séquence d'acides aminés</term><term>Virus du SRAS ()</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Membrane Glycoproteins</term><term>Peptides</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Membrane Glycoproteins</term><term>Peptides</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Peptides</term></keywords><keywords scheme="MESH" type="chemical" qualifier="physiology" xml:lang="en"><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Peptides</term><term>Protéines de l'enveloppe virale</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Peptides</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Cell Fusion</term><term>Cell Line</term><term>Humans</term><term>Molecular Sequence Data</term><term>Protein Structure, Tertiary</term><term>Sequence Alignment</term><term>Spike Glycoprotein, Coronavirus</term><term>Virus Internalization</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Alignement de séquences</term><term>Animaux</term><term>Données de séquences moléculaires</term><term>Fusion cellulaire</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires</term><term>Humains</term><term>Lignée cellulaire</term><term>Peptides</term><term>Protéines de l'enveloppe virale</term><term>Pénétration virale</term><term>Structure tertiaire des protéines</term><term>Séquence d'acides aminés</term><term>Virus du SRAS</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Many viral fusion proteins are primed by proteolytic cleavage near their fusion peptides. While the coronavirus (CoV) spike (S) protein is known to be cleaved at the S1/S2 boundary, this cleavage site is not closely linked to a fusion peptide. However, a second cleavage site has been identified in the severe acute respiratory syndrome CoV (SARS-CoV) S2 domain (R797). Here, we investigated whether this internal cleavage of S2 exposes a viral fusion peptide. We show that the residues immediately C-terminal to the SARS-CoV S2 cleavage site SFIEDLLFNKVTLADAGF are very highly conserved across all CoVs. Mutagenesis studies of these residues in SARS-CoV S, followed by cell-cell fusion and pseudotyped virion infectivity assays, showed a critical role for residues L803, L804, and F805 in membrane fusion. Mutation of the most N-terminal residue (S798) had little or no effect on membrane fusion. Biochemical analyses of synthetic peptides corresponding to the proposed S2 fusion peptide also showed an important role for this region in membrane fusion and indicated the presence of alpha-helical structure. We propose that proteolytic cleavage within S2 exposes a novel internal fusion peptide for SARS-CoV S, which may be conserved across the Coronaviridae.</div></front></TEI></pubmed></double></record>Pour manipuler ce document sous Unix (Dilib)
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