Second Site Mutations in the N-Terminus of the Major Capsid Protein (VP5) Overcome a Block at the Maturation Cleavage Site of the Capsid Scaffold Proteins of Herpes Simplex Virus Type 1
Identifieur interne : 003902 ( Main/Exploration ); précédent : 003901; suivant : 003903Second Site Mutations in the N-Terminus of the Major Capsid Protein (VP5) Overcome a Block at the Maturation Cleavage Site of the Capsid Scaffold Proteins of Herpes Simplex Virus Type 1
Auteurs : Prashant Desai ; Stanley PersonSource :
- Virology [ 0042-6822 ] ; 1999.
English descriptors
- Teeft :
- Amino, Amino acid residues, Amino acid substitution, Amino acid substitutions, Amino acids, Bacteriophage, Biol, Booy, Capsid, Capsid assembly, Capsid formation, Capsid maturation, Capsid proteins, Cleavage, Cleavage site, Codon, Desai, Ecori, Ecori site, Encoding, Enzyme activities, Enzyme activity, Gene encoding, Gene products, Herpes, Herpes simplex virus, Herpes simplex virus capsid, Herpes simplex virus type, Homa, Insect cells, Major capsid protein, Marker transfer, Maturation, Mutant, Mutant virus, Mutation, Newcomb, Nonpermissive, Nonpermissive cells, Open reading frame, Original mutation, Plaque, Plasmid, Protease, Protease cleavage, Protease cleavage site, Protease release site, Recombinant baculoviruses, Relative amount, Residue, Reversion, Revertant, Revertant viruses, Revertants, Rixon, Roizman, Scaffold, Scaffold maturation site, Scaffold molecules, Scaffold protein, Scaffold proteins, Second site, Second site mutations, Sequence analysis, Simplex, Small plaques, Southern blots, Spontaneous mutations, Steven, Sucrose gradients, Suppressor, Suppressor mutations, Tatman, Thomsen, Trus, Vero, Vero cell monolayers, Vero cells, Viral, Virol, Virology, Virus, Yeast system.
Abstract
Abstract: VP5, the major capsid protein of herpes simplex virus type 1 (HSV-1), interacts with the C-terminal residues of the scaffold molecules encoded by the overlapping UL26 and UL26.5 open reading frames. Scaffold molecules are cleaved by a UL26 encoded protease (VP24) as part of the normal capsid assembly process. In this study, residues of VP5 have been identified that alter its interaction with the C-terminal residues of the scaffold proteins. A previously isolated virus (KUL26-610/611) was used that encoded a lethal mutation in the UL26 and UL26.5 open reading frames and required a transformed cell line that expresses these proteins for virus growth. The scaffold maturation cleavage site between amino acids 610 and 611 was blocked by changing Ala-Ser to Glu-Phe, which generated a new EcoRI restriction site. Revertant viruses, that formed small plaques on nontransformed cells, were detected at a frequency of 1:3800. Nine revertants were isolated, and all of them retained the EcoRI site and therefore were due to mutations at a second site. The second site mutations were extragenic. Using marker-transfer techniques, the mutation in one of the revertants was mapped to the 5′ region of the gene encoding VP5. DNA sequence analysis was performed for the N-terminal 571 codons encoding VP5 for all of the revertant viruses. Six of the nine revertants showed a single base pair change that caused an amino acid substitution between residues 30 and 78 of VP5. Three of these were identical and changed Ala to Val at residue 78. The data provide a partial map of residues of VP5 that alter its interaction with scaffold proteins blocked at their normal cleavage site. The yeast two-hybrid system was used as a measure of the interaction between mutant VP5 and scaffold molecules and varied from 11% to nearly 100%, relative to wild-type VP5. One revertant gave no detectable interaction by this assay. The amount of UL26 encoded protease (VP24) in B capsids for KUL26-610/611 and for revertants was 7% and 25%, respectively, relative to the amount in capsids for wild-type virus. The lack of retention of the viral protease in the mutant virus and a fourfold increase for the revertants suggest an additional essential function for VP24 in capsid maturation, and a role in DNA packaging is indicated.
Url:
DOI: 10.1006/viro.1999.9877
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 002551
- to stream Istex, to step Curation: 002551
- to stream Istex, to step Checkpoint: 001155
- to stream Main, to step Merge: 003947
- to stream Main, to step Curation: 003902
Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Second Site Mutations in the N-Terminus of the Major Capsid Protein (VP5) Overcome a Block at the Maturation Cleavage Site of the Capsid Scaffold Proteins of Herpes Simplex Virus Type 1</title>
<author><name sortKey="Desai, Prashant" sort="Desai, Prashant" uniqKey="Desai P" first="Prashant" last="Desai">Prashant Desai</name>
</author>
<author><name sortKey="Person, Stanley" sort="Person, Stanley" uniqKey="Person S" first="Stanley" last="Person">Stanley Person</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">ISTEX</idno>
<idno type="RBID">ISTEX:7EACB6F7443A0AAECB414D23A2960A997FA21DBA</idno>
<date when="1999" year="1999">1999</date>
<idno type="doi">10.1006/viro.1999.9877</idno>
<idno type="url">https://api.istex.fr/ark:/67375/6H6-KJX0L6ZZ-9/fulltext.pdf</idno>
<idno type="wicri:Area/Istex/Corpus">002551</idno>
<idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">002551</idno>
<idno type="wicri:Area/Istex/Curation">002551</idno>
<idno type="wicri:Area/Istex/Checkpoint">001155</idno>
<idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001155</idno>
<idno type="wicri:doubleKey">0042-6822:1999:Desai P:second:site:mutations</idno>
<idno type="wicri:Area/Main/Merge">003947</idno>
<idno type="wicri:Area/Main/Curation">003902</idno>
<idno type="wicri:Area/Main/Exploration">003902</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Second Site Mutations in the N-Terminus of the Major Capsid Protein (VP5) Overcome a Block at the Maturation Cleavage Site of the Capsid Scaffold Proteins of Herpes Simplex Virus Type 1</title>
<author><name sortKey="Desai, Prashant" sort="Desai, Prashant" uniqKey="Desai P" first="Prashant" last="Desai">Prashant Desai</name>
<affiliation><wicri:noCountry code="subField">21205</wicri:noCountry>
</affiliation>
</author>
<author><name sortKey="Person, Stanley" sort="Person, Stanley" uniqKey="Person S" first="Stanley" last="Person">Stanley Person</name>
<affiliation><wicri:noCountry code="subField">21205</wicri:noCountry>
</affiliation>
</author>
</analytic>
<monogr></monogr>
<series><title level="j">Virology</title>
<title level="j" type="abbrev">YVIRO</title>
<idno type="ISSN">0042-6822</idno>
<imprint><publisher>ELSEVIER</publisher>
<date type="published" when="1999">1999</date>
<biblScope unit="volume">261</biblScope>
<biblScope unit="issue">2</biblScope>
<biblScope unit="page" from="357">357</biblScope>
<biblScope unit="page" to="366">366</biblScope>
</imprint>
<idno type="ISSN">0042-6822</idno>
</series>
</biblStruct>
</sourceDesc>
<seriesStmt><idno type="ISSN">0042-6822</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Amino</term>
<term>Amino acid residues</term>
<term>Amino acid substitution</term>
<term>Amino acid substitutions</term>
<term>Amino acids</term>
<term>Bacteriophage</term>
<term>Biol</term>
<term>Booy</term>
<term>Capsid</term>
<term>Capsid assembly</term>
<term>Capsid formation</term>
<term>Capsid maturation</term>
<term>Capsid proteins</term>
<term>Cleavage</term>
<term>Cleavage site</term>
<term>Codon</term>
<term>Desai</term>
<term>Ecori</term>
<term>Ecori site</term>
<term>Encoding</term>
<term>Enzyme activities</term>
<term>Enzyme activity</term>
<term>Gene encoding</term>
<term>Gene products</term>
<term>Herpes</term>
<term>Herpes simplex virus</term>
<term>Herpes simplex virus capsid</term>
<term>Herpes simplex virus type</term>
<term>Homa</term>
<term>Insect cells</term>
<term>Major capsid protein</term>
<term>Marker transfer</term>
<term>Maturation</term>
<term>Mutant</term>
<term>Mutant virus</term>
<term>Mutation</term>
<term>Newcomb</term>
<term>Nonpermissive</term>
<term>Nonpermissive cells</term>
<term>Open reading frame</term>
<term>Original mutation</term>
<term>Plaque</term>
<term>Plasmid</term>
<term>Protease</term>
<term>Protease cleavage</term>
<term>Protease cleavage site</term>
<term>Protease release site</term>
<term>Recombinant baculoviruses</term>
<term>Relative amount</term>
<term>Residue</term>
<term>Reversion</term>
<term>Revertant</term>
<term>Revertant viruses</term>
<term>Revertants</term>
<term>Rixon</term>
<term>Roizman</term>
<term>Scaffold</term>
<term>Scaffold maturation site</term>
<term>Scaffold molecules</term>
<term>Scaffold protein</term>
<term>Scaffold proteins</term>
<term>Second site</term>
<term>Second site mutations</term>
<term>Sequence analysis</term>
<term>Simplex</term>
<term>Small plaques</term>
<term>Southern blots</term>
<term>Spontaneous mutations</term>
<term>Steven</term>
<term>Sucrose gradients</term>
<term>Suppressor</term>
<term>Suppressor mutations</term>
<term>Tatman</term>
<term>Thomsen</term>
<term>Trus</term>
<term>Vero</term>
<term>Vero cell monolayers</term>
<term>Vero cells</term>
<term>Viral</term>
<term>Virol</term>
<term>Virology</term>
<term>Virus</term>
<term>Yeast system</term>
</keywords>
</textClass>
<langUsage><language ident="en">en</language>
</langUsage>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">Abstract: VP5, the major capsid protein of herpes simplex virus type 1 (HSV-1), interacts with the C-terminal residues of the scaffold molecules encoded by the overlapping UL26 and UL26.5 open reading frames. Scaffold molecules are cleaved by a UL26 encoded protease (VP24) as part of the normal capsid assembly process. In this study, residues of VP5 have been identified that alter its interaction with the C-terminal residues of the scaffold proteins. A previously isolated virus (KUL26-610/611) was used that encoded a lethal mutation in the UL26 and UL26.5 open reading frames and required a transformed cell line that expresses these proteins for virus growth. The scaffold maturation cleavage site between amino acids 610 and 611 was blocked by changing Ala-Ser to Glu-Phe, which generated a new EcoRI restriction site. Revertant viruses, that formed small plaques on nontransformed cells, were detected at a frequency of 1:3800. Nine revertants were isolated, and all of them retained the EcoRI site and therefore were due to mutations at a second site. The second site mutations were extragenic. Using marker-transfer techniques, the mutation in one of the revertants was mapped to the 5′ region of the gene encoding VP5. DNA sequence analysis was performed for the N-terminal 571 codons encoding VP5 for all of the revertant viruses. Six of the nine revertants showed a single base pair change that caused an amino acid substitution between residues 30 and 78 of VP5. Three of these were identical and changed Ala to Val at residue 78. The data provide a partial map of residues of VP5 that alter its interaction with scaffold proteins blocked at their normal cleavage site. The yeast two-hybrid system was used as a measure of the interaction between mutant VP5 and scaffold molecules and varied from 11% to nearly 100%, relative to wild-type VP5. One revertant gave no detectable interaction by this assay. The amount of UL26 encoded protease (VP24) in B capsids for KUL26-610/611 and for revertants was 7% and 25%, respectively, relative to the amount in capsids for wild-type virus. The lack of retention of the viral protease in the mutant virus and a fourfold increase for the revertants suggest an additional essential function for VP24 in capsid maturation, and a role in DNA packaging is indicated.</div>
</front>
</TEI>
<affiliations><list></list>
<tree><noCountry><name sortKey="Desai, Prashant" sort="Desai, Prashant" uniqKey="Desai P" first="Prashant" last="Desai">Prashant Desai</name>
<name sortKey="Person, Stanley" sort="Person, Stanley" uniqKey="Person S" first="Stanley" last="Person">Stanley Person</name>
</noCountry>
</tree>
</affiliations>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 003902 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 003902 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Sante |area= MersV1 |flux= Main |étape= Exploration |type= RBID |clé= ISTEX:7EACB6F7443A0AAECB414D23A2960A997FA21DBA |texte= Second Site Mutations in the N-Terminus of the Major Capsid Protein (VP5) Overcome a Block at the Maturation Cleavage Site of the Capsid Scaffold Proteins of Herpes Simplex Virus Type 1 }}
This area was generated with Dilib version V0.6.33. |