Detecting the Non-conventional mRNA Splicing and Translational Activation of HAC1 in Budding Yeast

Weihan Li, Robert H. Singer

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Protein-folding homeostasis in the endoplasmic reticulum (ER) is maintained by the unfolded protein response (UPR). UPR in Saccharomyces cerevisiae is regulated by a bZIP transcription factor, Hac1p. Under non-stress condition, HAC1 mRNA is translationally repressed. When un- or mis-folded proteins accumulate in the ER, HAC1 mRNA undergoes non-conventional mRNA splicing. The spliced HAC1 mRNA is translationally active and produces functional Hac1p, which initiates a transcriptional response that restores ER protein-folding homeostasis. Thus, the activation of yeast UPR is tightly regulated by HAC1 mRNA splicing. Here, we describe two methods that are used to monitor the splicing and translational status of HAC1 mRNA in budding yeast.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages113-120
Number of pages8
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2378
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Budding yeast
  • ER stress
  • HAC1
  • Non-conventional mRNA splicing
  • Unfolded protein response

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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