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Generation of catalytic human Ago4 identifies structural elements important for RNA cleavage.
Hauptmann, Judith; Kater, Lukas; Löffler, Patrick; Merkl, Rainer; Meister, Gunter.
Afiliación
  • Hauptmann J; Biochemistry Center Regensburg (BZR), Laboratory for RNA Biology, University of Regensburg, 93053 Regensburg, Germany.
  • Kater L; Biochemistry Center Regensburg (BZR), Laboratory for RNA Biology, University of Regensburg, 93053 Regensburg, Germany.
  • Löffler P; Institute of Biophysics and Physical Biochemistry, University of Regensburg, 93053 Regensburg, Germany.
  • Merkl R; Institute of Biophysics and Physical Biochemistry, University of Regensburg, 93053 Regensburg, Germany.
  • Meister G; Biochemistry Center Regensburg (BZR), Laboratory for RNA Biology, University of Regensburg, 93053 Regensburg, Germany gunter.meister@vkl.uni-regensburg.de.
RNA ; 20(10): 1532-8, 2014 Oct.
Article en En | MEDLINE | ID: mdl-25114291
Argonaute proteins bind small RNAs and mediate cleavage of complementary target RNAs. The human Argonaute protein Ago4 is catalytically inactive, although it is highly similar to catalytic Ago2. Here, we have generated Ago2-Ago4 chimeras and analyzed their cleavage activity in vitro. We identify several specific features that inactivate Ago4: the catalytic center, short sequence elements in the N-terminal domain, and an Ago4-specific insertion in the catalytic domain. In addition, we show that Ago2-mediated cleavage of the noncanonical miR-451 precursor can be carried out by any catalytic human Ago protein. Finally, phylogenetic analyses establish evolutionary distances between the Ago proteins. Interestingly, these distances do not fully correlate with the structural changes inactivating them, suggesting functional adaptations of individual human Ago proteins.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Factores Eucarióticos de Iniciación / Proteínas Argonautas / División del ARN Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: RNA Asunto de la revista: BIOLOGIA MOLECULAR Año: 2014 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Factores Eucarióticos de Iniciación / Proteínas Argonautas / División del ARN Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: RNA Asunto de la revista: BIOLOGIA MOLECULAR Año: 2014 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Estados Unidos