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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Intramural

MeSH terms

  • Genes, Reporter
  • HEK293 Cells
  • Heat Shock Transcription Factors / metabolism*
  • Heat-Shock Proteins / genetics
  • Heat-Shock Response*
  • Humans
  • Luciferases / genetics*
  • Molecular Biology / methods*
  • Transcriptional Activation*

Substances

  • HSF1 protein, human
  • Heat Shock Transcription Factors
  • Heat-Shock Proteins
  • Luciferases