Monitoring of the heat shock response with a real-time luciferase reporter

Toshiki Kijima, Takanori Eguchi, Len Neckers, Thomas L. Prince

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

The heat shock response (HSR) is a cellular mechanism for counteracting acute proteotoxic stress. In eukaryotes, transcriptional activation of the HSR is regulated by heat shock factor 1 (HSF1). Activation of HSF1 induces the expression of heat shock proteins (HSPs) that function as molecular chaperones to fold and maintain the three-dimensional structure of misfolded proteins. The regulation of the degree and duration of the HSR is controlled by multiple biochemical mechanisms that include posttranslational modification of HSF1 and numerous protein-protein interactions. In this chapter, we describe a method to evaluate the activation and deactivation of the HSR at the transcriptional level using a short half-life luciferase reporter assay. This assay can be used to further characterize the HSR or as a screen for small-molecule inducers, amplifiers, or repressors.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages35-45
Number of pages11
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1709
ISSN (Print)1064-3745

Keywords

  • Drug screen
  • Heat shock factor 1 (HSF1)
  • Heat shock protein 90 (HSP90)
  • Heat shock response
  • Luciferase assay
  • Real-time

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Kijima, T., Eguchi, T., Neckers, L., & Prince, T. L. (2018). Monitoring of the heat shock response with a real-time luciferase reporter. In Methods in Molecular Biology (pp. 35-45). (Methods in Molecular Biology; Vol. 1709). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7477-1_3