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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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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