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Analysis of the Heat Shock Factor Complex in Mammalian HSP70 Promoter

  • Protocol
Stress Responses

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1292))

Abstract

The heat shock response is characterized by the induction of heat shock proteins (HSPs) and is one of prominent mechanisms that regulate proteostasis capacity in the cell. In mammals, heat shock factor 1 (HSF1) regulates the expression of HSPs transcriptionally in both unstressed and stressed cells. Recent reports show that the HSF1-RPA complex constitutively gains access to nucleosomal DNA in part by recruiting a histone chaperone and a chromatin-remodeling component. Here, we describe the strategies to substitute endogenous HSF1 with ectopically expressed HSF1 or its mutant and to detect the occupancy of HSF1 transcription complex including RPA in vivo on two heat shock response elements located close together in the human or mouse HSP70 promoters by chromatin immunoprecipitation assay with high sensitivity and specificity.

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Acknowledgment

This work was supported in part by MEXT/JSPS KAKENHI Grant Number 3307, 24390081, 25430090, 25440010, the Takeda Science Foundation Special Project Research, the Uehara Memorial Foundation, and the Yamaguchi University Research Project on STRESS.

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Authors and Affiliations

  1. Departments of Biochemistry and Molecular Biology, Yamaguchi University School of Medicine, Minami-Kogushi 1-1-1, Ube, 755-8505, Japan

    Mitsuaki Fujimoto, Ryosuke Takii, Naoki Hayashida & Akira Nakai

Authors
  1. Mitsuaki Fujimoto
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  2. Ryosuke Takii
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  3. Naoki Hayashida
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  4. Akira Nakai
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Corresponding author

Correspondence to Akira Nakai .

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Editors and Affiliations

  1. Wentworth Institute of Technology, Boston, Massachusetts, USA

    Christine M. Oslowski

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Fujimoto, M., Takii, R., Hayashida, N., Nakai, A. (2015). Analysis of the Heat Shock Factor Complex in Mammalian HSP70 Promoter. In: Oslowski, C. (eds) Stress Responses. Methods in Molecular Biology, vol 1292. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2522-3_4

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  • DOI: https://doi.org/10.1007/978-1-4939-2522-3_4

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2521-6

  • Online ISBN: 978-1-4939-2522-3

  • eBook Packages: Springer Protocols

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