Identifying SARS-CoV membrane protein amino acid residues linked to virus-like particle assembly.
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 analy...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3659117?pdf=render |
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| author | Ying-Tzu Tseng Chia-Hui Chang Shiu-Mei Wang Kuo-Jung Huang Chin-Tien Wang |
| author_facet | Ying-Tzu Tseng Chia-Hui Chang Shiu-Mei Wang Kuo-Jung Huang Chin-Tien Wang |
| author_sort | Ying-Tzu Tseng |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-12T15:00:38Z |
| format | Article |
| id | doaj.art-2df30cde59684be581bb075747e97a13 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T15:00:38Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-2df30cde59684be581bb075747e97a132022-12-22T00:20:49ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0185e6401310.1371/journal.pone.0064013Identifying SARS-CoV membrane protein amino acid residues linked to virus-like particle assembly.Ying-Tzu TsengChia-Hui ChangShiu-Mei WangKuo-Jung HuangChin-Tien WangSevere 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.http://europepmc.org/articles/PMC3659117?pdf=render |
| spellingShingle | Ying-Tzu Tseng Chia-Hui Chang Shiu-Mei Wang Kuo-Jung Huang Chin-Tien Wang Identifying SARS-CoV membrane protein amino acid residues linked to virus-like particle assembly. PLoS ONE |
| title | Identifying SARS-CoV membrane protein amino acid residues linked to virus-like particle assembly. |
| title_full | Identifying SARS-CoV membrane protein amino acid residues linked to virus-like particle assembly. |
| title_fullStr | Identifying SARS-CoV membrane protein amino acid residues linked to virus-like particle assembly. |
| title_full_unstemmed | Identifying SARS-CoV membrane protein amino acid residues linked to virus-like particle assembly. |
| title_short | Identifying SARS-CoV membrane protein amino acid residues linked to virus-like particle assembly. |
| title_sort | identifying sars cov membrane protein amino acid residues linked to virus like particle assembly |
| url | http://europepmc.org/articles/PMC3659117?pdf=render |
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