Analysis of Constructed E Gene Mutants of Mouse Hepatitis Virus Confirms a Pivotal Role for E Protein in Coronavirus Assembly
Identifieur interne : 001956 ( Main/Exploration ); précédent : 001955; suivant : 001957Analysis of Constructed E Gene Mutants of Mouse Hepatitis Virus Confirms a Pivotal Role for E Protein in Coronavirus Assembly
Auteurs : Françoise Fischer ; Carola F. Stegen ; Paul S. Masters ; William A. SamsonoffSource :
- Journal of Virology [ 0022-538X ] ; 1998.
Abstract
Expression studies have shown that the coronavirus small envelope protein E and the much more abundant membrane glycoprotein M are both necessary and sufficient for the assembly of virus-like particles in cells. As a step toward understanding the function of the mouse hepatitis virus (MHV) E protein, we carried out clustered charged-to-alanine mutagenesis on the E gene and incorporated the resulting mutations into the MHV genome by targeted recombination. Of the four possible clustered charged-to-alanine E gene mutants, one was apparently lethal and one had a wild-type phenotype. The two other mutants were partially temperature sensitive, forming small plaques at the nonpermissive temperature. Revertant analyses of these two mutants demonstrated that the created mutations were responsible for the temperature-sensitive phenotype of each and provided support for possible interactions among E protein monomers. Both temperature-sensitive mutants were also found to be markedly thermolabile when grown at the permissive temperature, suggesting that there was a flaw in their assembly. Most significantly, when virions of one of the mutants were examined by electron microscopy, they were found to have strikingly aberrant morphology in comparison to the wild type: most mutant virions had pinched and elongated shapes that were rarely seen among wild-type virions. These results demonstrate an important, probably essential, role for the E protein in coronavirus morphogenesis.
Url:
PubMed: 9733825
PubMed Central: 110113
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Pmc, to step Corpus: 000155
- to stream Pmc, to step Curation: 000155
- to stream Pmc, to step Checkpoint: 000A38
- to stream Ncbi, to step Merge: 001691
- to stream Ncbi, to step Curation: 001691
- to stream Ncbi, to step Checkpoint: 001691
- to stream Main, to step Merge: 001969
- to stream Main, to step Curation: 001956
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Analysis of Constructed E Gene Mutants of Mouse Hepatitis Virus Confirms a Pivotal Role for E Protein in Coronavirus Assembly</title><author><name sortKey="Fischer, Francoise" sort="Fischer, Francoise" uniqKey="Fischer F" first="Françoise" last="Fischer">Françoise Fischer</name><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation></author><author><name sortKey="Stegen, Carola F" sort="Stegen, Carola F" uniqKey="Stegen C" first="Carola F." last="Stegen">Carola F. Stegen</name><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation></author><author><name sortKey="Masters, Paul S" sort="Masters, Paul S" uniqKey="Masters P" first="Paul S." last="Masters">Paul S. Masters</name><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation></author><author><name sortKey="Samsonoff, William A" sort="Samsonoff, William A" uniqKey="Samsonoff W" first="William A." last="Samsonoff">William A. Samsonoff</name><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">9733825</idno><idno type="pmc">110113</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC110113</idno><idno type="RBID">PMC:110113</idno><date when="1998">1998</date><idno type="wicri:Area/Pmc/Corpus">000155</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000155</idno><idno type="wicri:Area/Pmc/Curation">000155</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000155</idno><idno type="wicri:Area/Pmc/Checkpoint">000A38</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000A38</idno><idno type="wicri:Area/Ncbi/Merge">001691</idno><idno type="wicri:Area/Ncbi/Curation">001691</idno><idno type="wicri:Area/Ncbi/Checkpoint">001691</idno><idno type="wicri:doubleKey">0022-538X:1998:Fischer F:analysis:of:constructed</idno><idno type="wicri:Area/Main/Merge">001969</idno><idno type="wicri:Area/Main/Curation">001956</idno><idno type="wicri:Area/Main/Exploration">001956</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Analysis of Constructed E Gene Mutants of Mouse Hepatitis Virus Confirms a Pivotal Role for E Protein in Coronavirus Assembly</title><author><name sortKey="Fischer, Francoise" sort="Fischer, Francoise" uniqKey="Fischer F" first="Françoise" last="Fischer">Françoise Fischer</name><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation></author><author><name sortKey="Stegen, Carola F" sort="Stegen, Carola F" uniqKey="Stegen C" first="Carola F." last="Stegen">Carola F. Stegen</name><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation></author><author><name sortKey="Masters, Paul S" sort="Masters, Paul S" uniqKey="Masters P" first="Paul S." last="Masters">Paul S. Masters</name><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation></author><author><name sortKey="Samsonoff, William A" sort="Samsonoff, William A" uniqKey="Samsonoff W" first="William A." last="Samsonoff">William A. Samsonoff</name><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></affiliation><affiliation><nlm:aff id="N0xa04c090.0x9fb7ea8"></nlm:aff></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="1998">1998</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Expression studies have shown that the coronavirus small envelope protein E and the much more abundant membrane glycoprotein M are both necessary and sufficient for the assembly of virus-like particles in cells. As a step toward understanding the function of the mouse hepatitis virus (MHV) E protein, we carried out clustered charged-to-alanine mutagenesis on the E gene and incorporated the resulting mutations into the MHV genome by targeted recombination. Of the four possible clustered charged-to-alanine E gene mutants, one was apparently lethal and one had a wild-type phenotype. The two other mutants were partially temperature sensitive, forming small plaques at the nonpermissive temperature. Revertant analyses of these two mutants demonstrated that the created mutations were responsible for the temperature-sensitive phenotype of each and provided support for possible interactions among E protein monomers. Both temperature-sensitive mutants were also found to be markedly thermolabile when grown at the permissive temperature, suggesting that there was a flaw in their assembly. Most significantly, when virions of one of the mutants were examined by electron microscopy, they were found to have strikingly aberrant morphology in comparison to the wild type: most mutant virions had pinched and elongated shapes that were rarely seen among wild-type virions. These results demonstrate an important, probably essential, role for the E protein in coronavirus morphogenesis.</p></div></front></TEI><affiliations><list></list><tree><noCountry><name sortKey="Fischer, Francoise" sort="Fischer, Francoise" uniqKey="Fischer F" first="Françoise" last="Fischer">Françoise Fischer</name><name sortKey="Masters, Paul S" sort="Masters, Paul S" uniqKey="Masters P" first="Paul S." last="Masters">Paul S. Masters</name><name sortKey="Samsonoff, William A" sort="Samsonoff, William A" uniqKey="Samsonoff W" first="William A." last="Samsonoff">William A. Samsonoff</name><name sortKey="Stegen, Carola F" sort="Stegen, Carola F" uniqKey="Stegen C" first="Carola F." last="Stegen">Carola F. Stegen</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Sante/explor/CovidV2/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001956 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 001956 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Sante
|area= CovidV2
|flux= Main
|étape= Exploration
|type= RBID
|clé= PMC:110113
|texte= Analysis of Constructed E Gene Mutants of Mouse Hepatitis Virus Confirms a Pivotal Role for E Protein in Coronavirus Assembly
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:9733825" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a CovidV2
| This area was generated with Dilib version V0.6.33. |  |