Purification, molecular cloning, and cell-specific gene expression of the alkaloid-accumulation associated protein CrPS in Catharanthus roseus
Identifieur interne : 000674 ( Istex/Curation ); précédent : 000673; suivant : 000675Purification, molecular cloning, and cell-specific gene expression of the alkaloid-accumulation associated protein CrPS in Catharanthus roseus
Auteurs : Diane Leme Nager [France, Espagne] ; Lazhar Ouelhazi [France, Tunisie] ; Samira Mahroug [France] ; Bertrand Veau [France] ; Benoit St-Pierre [France] ; Marc Rideau [France] ; Jone Aguirreolea [Espagne] ; Vincent Burlat [France] ; Marc Clastre [France]Source :
- Journal of Experimental Botany [ 0022-0957 ] ; 2005-04.
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Abstract
Identification of molecular markers of monoterpenoid indole alkaloid (MIA) accumulation in cell-suspension cultures of Madagascar periwinkle (Catharanthus roseus (L.) G. Don) was performed by two-dimensional polyacrylamide gel electrophoresis. Comparison of the protein patterns from alkaloid-producing and non-producing cells showed the specific occurrence of a 28 kDa polypeptide restricted to cells accumulating MIAs. The polypeptide was purified by preparative two-dimensional gel electrophoresis, digested with trypsin, and microsequenced by the Edman degradation method. Cloning of the corresponding cDNA revealed that the protein which has been named CrPS (Catharanthus roseus Protein S) is a member of the α/β hydrolase superfamily. Time-course expression studies by northern blot analysis confirmed that CrPS gene expression was associated with MIA accumulation in cell suspension cultures. In the whole plant, multicellular compartmentation is required for alkaloid biosynthesis. In situ mRNA hybridization on developing leaves revealed that CrPS mRNA and transcripts encoding the first enzymes of the MIA pathway were co-localized in internal phloem parenchyma cells. The possible implication of the alkaloid-accumulation associated protein CrPS in the signal transduction pathway leading to MIA production is discussed.
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DOI: 10.1093/jxb/eri116
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Biotechnologies Végétales, Université de Tours, 31 Avenue Monge, F-37200 Tours, France</mods:affiliation><country xml:lang="fr">France</country><wicri:regionArea>EA2106 Biomolécules et Biotechnologies Végétales, Université de Tours, 31 Avenue Monge, F-37200 Tours</wicri:regionArea></affiliation></author><author><name sortKey="Veau, Bertrand" sort="Veau, Bertrand" uniqKey="Veau B" first="Bertrand" last="Veau">Bertrand Veau</name><affiliation wicri:level="1"><mods:affiliation>EA2106 Biomolécules et Biotechnologies Végétales, Université de Tours, 31 Avenue Monge, F-37200 Tours, France</mods:affiliation><country xml:lang="fr">France</country><wicri:regionArea>EA2106 Biomolécules et Biotechnologies Végétales, Université de Tours, 31 Avenue Monge, F-37200 Tours</wicri:regionArea></affiliation></author><author><name sortKey="St Pierre, Benoit" sort="St Pierre, Benoit" uniqKey="St Pierre B" first="Benoit" last="St-Pierre">Benoit St-Pierre</name><affiliation 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Aguirreolea</name><affiliation wicri:level="1"><mods:affiliation>Departamento de Fisiologia Vegetal, Universidad de Navarra, c/Irunlarrea s/n, 31008 Pamplona, Spain</mods:affiliation><country xml:lang="fr">Espagne</country><wicri:regionArea>Departamento de Fisiologia Vegetal, Universidad de Navarra, c/Irunlarrea s/n, 31008 Pamplona</wicri:regionArea></affiliation></author><author><name sortKey="Burlat, Vincent" sort="Burlat, Vincent" uniqKey="Burlat V" first="Vincent" last="Burlat">Vincent Burlat</name><affiliation wicri:level="1"><mods:affiliation>EA2106 Biomolécules et Biotechnologies Végétales, Université de Tours, 31 Avenue Monge, F-37200 Tours, France</mods:affiliation><country xml:lang="fr">France</country><wicri:regionArea>EA2106 Biomolécules et Biotechnologies Végétales, Université de Tours, 31 Avenue Monge, F-37200 Tours</wicri:regionArea></affiliation></author><author><name sortKey="Clastre, Marc" sort="Clastre, Marc" uniqKey="Clastre M" first="Marc" last="Clastre">Marc Clastre</name><affiliation wicri:level="1"><mods:affiliation>EA2106 Biomolécules et Biotechnologies Végétales, Université de Tours, 31 Avenue Monge, F-37200 Tours, France</mods:affiliation><country xml:lang="fr">France</country><wicri:regionArea>EA2106 Biomolécules et Biotechnologies Végétales, Université de Tours, 31 Avenue Monge, F-37200 Tours</wicri:regionArea></affiliation><affiliation><mods:affiliation>‡To whom correspondence should be addressed at Laboratoire de Biologie Moléculaire Végétale, Faculté des Sciences Pharmaceutiques, 31 Avenue Monge, F-37200 Tours, France. Fax: +33 247 27 66 60.</mods:affiliation></affiliation><affiliation wicri:level="1"><mods:affiliation>E-mail: marc.clastre@univ-tours.fr</mods:affiliation><country wicri:rule="url">France</country></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Experimental Botany</title><title level="j" type="abbrev">J. Exp. Bot.</title><idno type="ISSN">0022-0957</idno><idno type="eISSN">1460-2431</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="2005-04">2005-04</date><biblScope unit="volume">56</biblScope><biblScope unit="issue">414</biblScope><biblScope unit="page" from="1221">1221</biblScope><biblScope unit="page" to="1228">1228</biblScope></imprint><idno type="ISSN">0022-0957</idno></series><idno type="istex">F53C5249FD225A76AAD57081DD49169AA5A01B8F</idno><idno type="DOI">10.1093/jxb/eri116</idno><idno type="local">eri116</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0022-0957</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Catharanthus roseus</term><term>in situ RNA hybridization</term><term>monoterpenoid indole alkaloid</term><term>phytohormone</term><term>two-dimensional gel electrophoresis</term><term>α/β hydrolase</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Identification of molecular markers of monoterpenoid indole alkaloid (MIA) accumulation in cell-suspension cultures of Madagascar periwinkle (Catharanthus roseus (L.) G. Don) was performed by two-dimensional polyacrylamide gel electrophoresis. Comparison of the protein patterns from alkaloid-producing and non-producing cells showed the specific occurrence of a 28 kDa polypeptide restricted to cells accumulating MIAs. The polypeptide was purified by preparative two-dimensional gel electrophoresis, digested with trypsin, and microsequenced by the Edman degradation method. Cloning of the corresponding cDNA revealed that the protein which has been named CrPS (Catharanthus roseus Protein S) is a member of the α/β hydrolase superfamily. Time-course expression studies by northern blot analysis confirmed that CrPS gene expression was associated with MIA accumulation in cell suspension cultures. In the whole plant, multicellular compartmentation is required for alkaloid biosynthesis. In situ mRNA hybridization on developing leaves revealed that CrPS mRNA and transcripts encoding the first enzymes of the MIA pathway were co-localized in internal phloem parenchyma cells. The possible implication of the alkaloid-accumulation associated protein CrPS in the signal transduction pathway leading to MIA production is discussed.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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