Identifying SARS-CoV membrane protein amino acid residues linked to virus-like particle assembly.
Identifieur interne : 001919 ( Main/Exploration ); précédent : 001918; suivant : 001920Identifying SARS-CoV membrane protein amino acid residues linked to virus-like particle assembly.
Auteurs : Ying-Tzu Tseng [Taïwan] ; Chia-Hui Chang ; Shiu-Mei Wang ; Kuo-Jung Huang ; Chin-Tien WangSource :
- PloS one [ 1932-6203 ] ; 2013.
Descripteurs français
- KwdFr :
- Assemblage viral, Cellules HEK293, Cellules HeLa, Humains, Motifs d'acides aminés, Multimérisation de protéines, Mutagenèse dirigée, Protéines de la matrice virale (), Protéines de la matrice virale (génétique), Protéines de la matrice virale (métabolisme), Structure tertiaire des protéines, Substitution d'acide aminé, Virion (physiologie), Virus du SRAS (physiologie).
- MESH :
- génétique : Protéines de la matrice virale.
- métabolisme : Protéines de la matrice virale.
- physiologie : Virion, Virus du SRAS.
- Assemblage viral, Cellules HEK293, Cellules HeLa, Humains, Motifs d'acides aminés, Multimérisation de protéines, Mutagenèse dirigée, Protéines de la matrice virale, Structure tertiaire des protéines, Substitution d'acide aminé.
English descriptors
- KwdEn :
- Amino Acid Motifs, Amino Acid Substitution, HEK293 Cells, HeLa Cells, Humans, Mutagenesis, Site-Directed, Protein Multimerization, Protein Structure, Tertiary, SARS Virus (physiology), Viral Matrix Proteins (chemistry), Viral Matrix Proteins (genetics), Viral Matrix Proteins (metabolism), Virion (physiology), Virus Assembly.
- MESH :
- chemical , chemistry : Viral Matrix Proteins.
- chemical , genetics : Viral Matrix Proteins.
- chemical , metabolism : Viral Matrix Proteins.
- physiology : SARS Virus, Virion.
- Amino Acid Motifs, Amino Acid Substitution, HEK293 Cells, HeLa Cells, Humans, Mutagenesis, Site-Directed, Protein Multimerization, Protein Structure, Tertiary, Virus Assembly.
Abstract
Severe acute respiratory syndrome coronavirus (SARS-CoV) membrane (M) proteins are capable of self-assembly and release in the form of membrane-enveloped vesicles, and of forming virus-like particles (VLPs) when coexpressed with SARS-CoV nucleocapsid (N) protein. According to previous deletion analyses, M self-assembly involves multiple M sequence regions. To identify important M amino acid residues for VLP assembly, we coexpressed N with multiple M mutants containing substitution mutations at the amino-terminal ectodomain, carboxyl-terminal endodomain, or transmembrane segments. Our results indicate that a dileucine motif in the endodomain tail (218LL219) is required for efficient N packaging into VLPs. Results from cross-linking VLP analyses suggest that the cysteine residues 63, 85 and 158 are not in close proximity to the M dimer interface. We noted a significant reduction in M secretion due to serine replacement for C158, but not for C63 or C85. Further analysis suggests that C158 is involved in M-N interaction. In addition to mutations of the highly conserved 107-SWWSFNPE-114 motif, substitutions at codons W19, W57, P58, W91, Y94 or F95 all resulted in significantly reduced VLP yields, largely due to defective M secretion. VLP production was not significantly affected by a tryptophan replacement of Y94 or F95 or a phenylalanine replacement of W19, W57 or W91. Combined, these results indicate the involvement of specific M amino acids during SARS-CoV virus assembly, and suggest that aromatic residue retention at specific positions is critical for M function in terms of directing virus assembly.
DOI: 10.1371/journal.pone.0064013
PubMed: 23700447
Affiliations:
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Le document en format XML
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Chia Hui" uniqKey="Chang C" first="Chia-Hui" last="Chang">Chia-Hui Chang</name></author><author><name sortKey="Wang, Shiu Mei" sort="Wang, Shiu Mei" uniqKey="Wang S" first="Shiu-Mei" last="Wang">Shiu-Mei Wang</name></author><author><name sortKey="Huang, Kuo Jung" sort="Huang, Kuo Jung" uniqKey="Huang K" first="Kuo-Jung" last="Huang">Kuo-Jung Huang</name></author><author><name sortKey="Wang, Chin Tien" sort="Wang, Chin Tien" uniqKey="Wang C" first="Chin-Tien" last="Wang">Chin-Tien Wang</name></author></analytic><series><title level="j">PloS one</title><idno type="eISSN">1932-6203</idno><imprint><date when="2013" type="published">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Motifs</term><term>Amino Acid Substitution</term><term>HEK293 Cells</term><term>HeLa Cells</term><term>Humans</term><term>Mutagenesis, Site-Directed</term><term>Protein Multimerization</term><term>Protein Structure, Tertiary</term><term>SARS Virus (physiology)</term><term>Viral Matrix Proteins (chemistry)</term><term>Viral Matrix Proteins (genetics)</term><term>Viral Matrix Proteins (metabolism)</term><term>Virion (physiology)</term><term>Virus Assembly</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Assemblage viral</term><term>Cellules HEK293</term><term>Cellules HeLa</term><term>Humains</term><term>Motifs d'acides aminés</term><term>Multimérisation de protéines</term><term>Mutagenèse dirigée</term><term>Protéines de la matrice virale ()</term><term>Protéines de la matrice virale (génétique)</term><term>Protéines de la matrice virale (métabolisme)</term><term>Structure tertiaire des protéines</term><term>Substitution d'acide aminé</term><term>Virion (physiologie)</term><term>Virus du SRAS (physiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines de la matrice virale</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Protéines de la matrice virale</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Virion</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>SARS Virus</term><term>Virion</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Motifs</term><term>Amino Acid Substitution</term><term>HEK293 Cells</term><term>HeLa Cells</term><term>Humans</term><term>Mutagenesis, Site-Directed</term><term>Protein Multimerization</term><term>Protein Structure, Tertiary</term><term>Virus Assembly</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Assemblage viral</term><term>Cellules HEK293</term><term>Cellules HeLa</term><term>Humains</term><term>Motifs d'acides aminés</term><term>Multimérisation de protéines</term><term>Mutagenèse dirigée</term><term>Protéines de la matrice virale</term><term>Structure tertiaire des protéines</term><term>Substitution d'acide aminé</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome coronavirus (SARS-CoV) membrane (M) proteins are capable of self-assembly and release in the form of membrane-enveloped vesicles, and of forming virus-like particles (VLPs) when coexpressed with SARS-CoV nucleocapsid (N) protein. According to previous deletion analyses, M self-assembly involves multiple M sequence regions. To identify important M amino acid residues for VLP assembly, we coexpressed N with multiple M mutants containing substitution mutations at the amino-terminal ectodomain, carboxyl-terminal endodomain, or transmembrane segments. Our results indicate that a dileucine motif in the endodomain tail (218LL219) is required for efficient N packaging into VLPs. Results from cross-linking VLP analyses suggest that the cysteine residues 63, 85 and 158 are not in close proximity to the M dimer interface. We noted a significant reduction in M secretion due to serine replacement for C158, but not for C63 or C85. Further analysis suggests that C158 is involved in M-N interaction. In addition to mutations of the highly conserved 107-SWWSFNPE-114 motif, substitutions at codons W19, W57, P58, W91, Y94 or F95 all resulted in significantly reduced VLP yields, largely due to defective M secretion. VLP production was not significantly affected by a tryptophan replacement of Y94 or F95 or a phenylalanine replacement of W19, W57 or W91. Combined, these results indicate the involvement of specific M amino acids during SARS-CoV virus assembly, and suggest that aromatic residue retention at specific positions is critical for M function in terms of directing virus assembly.</div></front></TEI><affiliations><list><country><li>Taïwan</li></country></list><tree><noCountry><name sortKey="Chang, Chia Hui" sort="Chang, Chia Hui" uniqKey="Chang C" first="Chia-Hui" last="Chang">Chia-Hui Chang</name><name sortKey="Huang, Kuo Jung" sort="Huang, Kuo Jung" uniqKey="Huang K" first="Kuo-Jung" last="Huang">Kuo-Jung Huang</name><name sortKey="Wang, Chin Tien" sort="Wang, Chin Tien" uniqKey="Wang C" first="Chin-Tien" last="Wang">Chin-Tien Wang</name><name sortKey="Wang, Shiu Mei" sort="Wang, Shiu Mei" uniqKey="Wang S" first="Shiu-Mei" last="Wang">Shiu-Mei Wang</name></noCountry><country name="Taïwan"><noRegion><name sortKey="Tseng, Ying Tzu" sort="Tseng, Ying Tzu" uniqKey="Tseng Y" first="Ying-Tzu" last="Tseng">Ying-Tzu Tseng</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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