Cloning and sequencing of caspase 6 in rainbow trout, Oncorhynchus mykiss , and analysis of its expression under conditions known to induce apoptosis
Identifieur interne : 000623 ( Main/Exploration ); précédent : 000622; suivant : 000624Cloning and sequencing of caspase 6 in rainbow trout, Oncorhynchus mykiss , and analysis of its expression under conditions known to induce apoptosis
Auteurs : K. J. Laing [Royaume-Uni] ; J. Holland [Royaume-Uni] ; S. Bonilla [Royaume-Uni, Espagne] ; C. Cunningham [Norvège] ; C. J. Secombes [Royaume-Uni]Source :
- Developmental and Comparative Immunology [ 0145-305X ] ; 2000.
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Abstract
The rainbow trout caspase 6 gene has been cloned and sequenced. The open reading frame consisted of 906bp, which translated into a protein of 302 amino acids, containing the caspase active site pentapeptide (QACRG) and the caspase family signature (HADADCFVCVFLSHG). Amino acids involved in catalysis and those known to form the P1 carbohydrate binding pocket were conserved. Phylogenetic tree analysis showed a tight grouping with other known caspase 6 genes. Conserved aspartic acid residues at positions 33, 191 and 202 suggested that this molecule is produced as a proenzyme that is subsequently cleaved to release active subunits, with the region between Asp-191 and Ala-203 acting as a linker that is cleaved out. RT-PCR analysis revealed that the trout caspase 6 gene was expressed in brain, blood, gill, liver, head kidney and spleen. Addition of LPS or cortisol to head kidney leucocyte cultures had no effect upon caspase 6 expression. However, addition of LPS after preincubation with cortisol increased expression relative to control cultures. Incubation with RU486 abrogated this effect, confirming it was mediated via glucocorticoid receptors. Lastly, a confinement stress in vivo increased caspase 6 expression. The data are discussed with respect to the immunoregulatory role of apoptosis in fish immune responses.
Url:
DOI: 10.1016/S0145-305X(00)00061-6
Affiliations:
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J." last="Laing">K. J. Laing</name><affiliation wicri:level="1"><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Department of Zoology, University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ</wicri:regionArea><wicri:noRegion>Aberdeen AB24 2TZ</wicri:noRegion></affiliation><affiliation><wicri:noCountry code="no comma">1 Equal contribution to the work.</wicri:noCountry></affiliation></author><author><name sortKey="Holland, J" sort="Holland, J" uniqKey="Holland J" first="J." last="Holland">J. Holland</name><affiliation wicri:level="1"><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Department of Zoology, University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ</wicri:regionArea><wicri:noRegion>Aberdeen AB24 2TZ</wicri:noRegion></affiliation><affiliation><wicri:noCountry code="no comma">1 Equal contribution to the work.</wicri:noCountry></affiliation></author><author><name sortKey="Bonilla, S" sort="Bonilla, S" uniqKey="Bonilla S" first="S." last="Bonilla">S. Bonilla</name><affiliation wicri:level="1"><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Department of Zoology, University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ</wicri:regionArea><wicri:noRegion>Aberdeen AB24 2TZ</wicri:noRegion></affiliation><affiliation wicri:level="2"><country xml:lang="fr" wicri:curation="lc">Espagne</country><wicri:regionArea>2 Present address: Department of Cell Biology, University of Murcia, 30100 Murcia</wicri:regionArea><placeName><region nuts="2" type="communauté">Région de Murcie</region></placeName></affiliation></author><author><name sortKey="Cunningham, C" sort="Cunningham, C" uniqKey="Cunningham C" first="C." last="Cunningham">C. Cunningham</name><affiliation wicri:level="1"><country xml:lang="fr">Norvège</country><wicri:regionArea>SARS International Center for Marine Molecular Biology, 5008 Bergen</wicri:regionArea><wicri:noRegion>5008 Bergen</wicri:noRegion></affiliation></author><author><name sortKey="Secombes, C J" sort="Secombes, C J" uniqKey="Secombes C" first="C. J." last="Secombes">C. J. Secombes</name><affiliation wicri:level="1"><country xml:lang="fr">Royaume-Uni</country><wicri:regionArea>Department of Zoology, University of Aberdeen, Tillydrone Avenue, Aberdeen AB24 2TZ</wicri:regionArea><wicri:noRegion>Aberdeen AB24 2TZ</wicri:noRegion></affiliation><affiliation><wicri:noCountry code="no comma">Corresponding author. Tel.: +44-1224-272872; fax: +44-1224-272396</wicri:noCountry></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Royaume-Uni</country></affiliation></author></analytic><monogr></monogr><series><title level="j">Developmental and Comparative Immunology</title><title level="j" type="abbrev">DCI</title><idno type="ISSN">0145-305X</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="2000">2000</date><biblScope unit="volume">25</biblScope><biblScope unit="issue">4</biblScope><biblScope unit="page" from="303">303</biblScope><biblScope unit="page" to="312">312</biblScope></imprint><idno type="ISSN">0145-305X</idno></series><idno type="istex">B557C68F569F0AD361DAD61798D70DC248EEE113</idno><idno type="DOI">10.1016/S0145-305X(00)00061-6</idno><idno type="PII">S0145-305X(00)00061-6</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0145-305X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Caspase 6</term><term>Cortisol</term><term>Expression</term><term>LPS, lipopolysaccharide</term><term>ORF, open reading frame</term><term>PCR, polymerase chain reaction</term><term>RT-PCR</term><term>Rainbow trout</term><term>Stress</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The rainbow trout caspase 6 gene has been cloned and sequenced. The open reading frame consisted of 906bp, which translated into a protein of 302 amino acids, containing the caspase active site pentapeptide (QACRG) and the caspase family signature (HADADCFVCVFLSHG). Amino acids involved in catalysis and those known to form the P1 carbohydrate binding pocket were conserved. Phylogenetic tree analysis showed a tight grouping with other known caspase 6 genes. Conserved aspartic acid residues at positions 33, 191 and 202 suggested that this molecule is produced as a proenzyme that is subsequently cleaved to release active subunits, with the region between Asp-191 and Ala-203 acting as a linker that is cleaved out. RT-PCR analysis revealed that the trout caspase 6 gene was expressed in brain, blood, gill, liver, head kidney and spleen. Addition of LPS or cortisol to head kidney leucocyte cultures had no effect upon caspase 6 expression. However, addition of LPS after preincubation with cortisol increased expression relative to control cultures. Incubation with RU486 abrogated this effect, confirming it was mediated via glucocorticoid receptors. Lastly, a confinement stress in vivo increased caspase 6 expression. The data are discussed with respect to the immunoregulatory role of apoptosis in fish immune responses.</div></front></TEI><affiliations><list><country><li>Espagne</li><li>Norvège</li><li>Royaume-Uni</li></country><region><li>Région de Murcie</li></region></list><tree><country name="Royaume-Uni"><noRegion><name sortKey="Laing, K J" sort="Laing, K J" uniqKey="Laing K" first="K. J." last="Laing">K. J. Laing</name></noRegion><name sortKey="Bonilla, S" sort="Bonilla, S" uniqKey="Bonilla S" first="S." last="Bonilla">S. Bonilla</name><name sortKey="Holland, J" sort="Holland, J" uniqKey="Holland J" first="J." last="Holland">J. Holland</name><name sortKey="Secombes, C J" sort="Secombes, C J" uniqKey="Secombes C" first="C. J." last="Secombes">C. J. Secombes</name><name sortKey="Secombes, C J" sort="Secombes, C J" uniqKey="Secombes C" first="C. J." last="Secombes">C. J. Secombes</name></country><country name="Espagne"><region name="Région de Murcie"><name sortKey="Bonilla, S" sort="Bonilla, S" uniqKey="Bonilla S" first="S." last="Bonilla">S. Bonilla</name></region></country><country name="Norvège"><noRegion><name sortKey="Cunningham, C" sort="Cunningham, C" uniqKey="Cunningham C" first="C." last="Cunningham">C. Cunningham</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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