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Determining if an mRNA is a Substrate of Nonsense-Mediated mRNA Decay in Saccharomyces cerevisiae

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Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1507))

Abstract

Nonsense-mediated mRNA decay (NMD) is a conserved eukaryotic quality control mechanism which triggers decay of mRNAs harboring premature translation termination codons. In this chapter, I describe methods for monitoring the influence of NMD on mRNA abundance and decay rates in Saccharomyces cerevisiae. The descriptions include detailed methods for growing yeast cells, total RNA isolation, and Northern blotting. Although the chapter focuses on NMD, the methods can be easily adapted to assess the effect of other mRNA decay pathways.

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Acknowledgments

I thank Yang Zhou, Nathan Kenney, and Tracy Nissan for the helpful comments on the manuscript. The work in my laboratory is supported by Carl Tryggers Foundation (CTS13:206), Åke Wibergs Foundation (M14-0207), and Insamlingsstiftelsen Umeå universitet.

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Authors and Affiliations

  1. Department of Molecular Biology, Umeå University, 901 87, Umeå, Sweden

    Marcus J. O. Johansson

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  1. Marcus J. O. Johansson
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Correspondence to Marcus J. O. Johansson .

Editor information

Editors and Affiliations

  1. Department of Pathology, Yale University School of Medicine, New Haven, Connecticut, USA

    Narendra Wajapeyee

  2. Department of Pathology, Yale University School of Medicine, New Haven, Connecticut, USA

    Romi Gupta

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Johansson, M.J.O. (2017). Determining if an mRNA is a Substrate of Nonsense-Mediated mRNA Decay in Saccharomyces cerevisiae . In: Wajapeyee, N., Gupta, R. (eds) Eukaryotic Transcriptional and Post-Transcriptional Gene Expression Regulation. Methods in Molecular Biology, vol 1507. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6518-2_13

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  • DOI: https://doi.org/10.1007/978-1-4939-6518-2_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6516-8

  • Online ISBN: 978-1-4939-6518-2

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