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Methods Mol Biol. 2018;1709:35-45. doi: 10.1007/978-1-4939-7477-1_3.

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

Author information

1
Department of Urology, Tokyo Medical and Dental University Graduate School, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan.
2
Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, 700-8525, Japan.
3
Urologic Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
4
Department of Urology, Weis Center for Research, Geisinger Clinic, Danville, PA, 17822, USA. tprince@geisinger.edu.

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.

KEYWORDS:

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

PMID:
29177649
DOI:
10.1007/978-1-4939-7477-1_3
[Indexed for MEDLINE]

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