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Detection of HAC1 mRNA Splicing by RT-PCR in Saccharomyces cerevisiae

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The Unfolded Protein Response

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2378))

Abstract

HAC1 mRNA remains translationally repressed in the cytoplasm of the budding yeast Saccharomyces cerevisiae. Under conditions of cellular stress, a dual kinase RNase IRE1 (Inositol Requiring Enzyme-1) cleaves out an intervening sequence from the HAC1 mRNA. Cleaved mRNAs are then ligated by tRNA ligase, thus generating a spliced mRNA that translates an active transcription factor. This unconventional splicing of HAC1 mRNA in the cytoplasm is a molecular marker for various cellular stresses including oxidative stress and endoplasmic reticulum (ER) stress. This article describes a PCR-based protocol to detect the HAC1 mRNA splicing.

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Acknowledgment

This work was supported by a grant to M.D. from the U.S. National Institutes of Health (1R01GM124183).

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Authors and Affiliations

  1. Department of Biological Sciences, University of Wisconsin-Milwaukee, Milwaukee, WI, USA

    Jagadeesh Kumar Uppala & Madhusudan Dey

Authors
  1. Jagadeesh Kumar Uppala
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  2. Madhusudan Dey
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Corresponding author

Correspondence to Madhusudan Dey .

Editor information

Editors and Affiliations

  1. Instituto de Agroquímica y Tecnología de Alimentos, CSIC, Paterna, Spain

    Roberto Pérez-Torrado

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Uppala, J.K., Dey, M. (2022). Detection of HAC1 mRNA Splicing by RT-PCR in Saccharomyces cerevisiae . In: Pérez-Torrado, R. (eds) The Unfolded Protein Response. Methods in Molecular Biology, vol 2378. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1732-8_7

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  • DOI: https://doi.org/10.1007/978-1-0716-1732-8_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1731-1

  • Online ISBN: 978-1-0716-1732-8

  • eBook Packages: Springer Protocols

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