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Monitoring of the Heat Shock Response with a Real-Time Luciferase Reporter

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Chaperones

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2693))

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.

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Author information

Authors and Affiliations

  1. Geisinger Clinic, Danville, PA, USA

    Andrew Ackerman

  2. Department of Urology, Dokkyo Medical University, Tochigi, Japan

    Toshiki Kijima

  3. Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan

    Takanori Eguchi

  4. Ranok Therapeutics, Danville, PA, USA

    Thomas L. Prince

Authors
  1. Andrew Ackerman
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  2. Toshiki Kijima
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  3. Takanori Eguchi
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  4. Thomas L. Prince
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Corresponding author

Correspondence to Thomas L. Prince .

Editor information

Editors and Affiliations

  1. Harvard Medical School, Beth Israel Deaconess Medical Center, Boston, MA, USA

    Stuart K. Calderwood

  2. Danville, PA, USA

    Thomas L. Prince

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Ackerman, A., Kijima, T., Eguchi, T., Prince, T.L. (2023). Monitoring of the Heat Shock Response with a Real-Time Luciferase Reporter. In: Calderwood, S.K., Prince, T.L. (eds) Chaperones. Methods in Molecular Biology, vol 2693. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3342-7_1

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  • DOI: https://doi.org/10.1007/978-1-0716-3342-7_1

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3341-0

  • Online ISBN: 978-1-0716-3342-7

  • eBook Packages: Springer Protocols

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