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Modification of Selenoprotein mRNAs by Cap Tri-methylation

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Selenoproteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1661))

Abstract

Several selenoprotein mRNAs undergo 5′ cap maturation events whereby their classical monomethylated m7G cap becomes trimethylated (m3 2,2,7G) by the trimethylguanosine synthase 1 (Tgs1). Here, we describe immunoprecipitation methods for the detection of endogenous m3 2,2,7G-capped selenoprotein mRNAs from total cell extracts or after polysome fractionation of cytoplasmic extracts. We have also developed a method for the in vitro cap hypermethylation of selenoprotein mRNA transcripts using purified Tgs1 enzyme.

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Acknowledgments

We thank Thomas Monecke and Ralf Ficner for the gift of pGEX-6P hTgs1618-853 plasmid, Rémy Bordonné for the gift of pGEX-sTgs1577-851 plasmid.

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Authors and Affiliations

  1. Architecture et Réactivité de l’ARN, Centre National de la Recherche Scientifique, Institut de Biologie Moléculaire et Cellulaire, Université de Strasbourg, Strasbourg, 67000, France

    Anne-Sophie Gribling-Burrer, Gilbert Eriani & Christine Allmang

  2. Faculty of Medicine, ZBMZ, Institute of Biochemistry and Molecular Biology, University of Freiburg, Freiburg, 79104, Germany

    Anne-Sophie Gribling-Burrer

Authors
  1. Anne-Sophie Gribling-Burrer
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  2. Gilbert Eriani
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  3. Christine Allmang
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Corresponding author

Correspondence to Christine Allmang .

Editor information

Editors and Affiliations

  1. Centre International de Recherche en Infectiologie, CIRI, Lyon, France

    Laurent Chavatte

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Gribling-Burrer, AS., Eriani, G., Allmang, C. (2018). Modification of Selenoprotein mRNAs by Cap Tri-methylation. In: Chavatte, L. (eds) Selenoproteins. Methods in Molecular Biology, vol 1661. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7258-6_9

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  • DOI: https://doi.org/10.1007/978-1-4939-7258-6_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7257-9

  • Online ISBN: 978-1-4939-7258-6

  • eBook Packages: Springer Protocols

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