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A delayed antioxidant response in heat-stressed cells expressing a non-DNA binding HSF1 mutant

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Cell Stress and Chaperones Aims and scope

Abstract

To assess the consequences of inactivation of heat shock factor 1 (HSF1) during aging, we analyzed the effect of HSF1 K80Q, a mutant unable to bind DNA, and of dnHSF1, a mutant lacking the activation domain, on the transcriptome of cells 6 and 24 h after heat shock. The primary response to heat shock (6 h recovery), of which 30 % was HSF1-dependent, had decayed 24 h after heat shock in control cells but was extended in HSF1 K80Q and dnHSF1 cells. Comparison with literature data showed that even the HSF1 dependent primary stress response is largely cell specific. HSF1 K80Q, but not HSF1 siRNA-treated, cells showed a delayed stress response: an increase in transcript levels of HSF1 target genes 24 h after heat stress. Knockdown of NRF2, but not of ATF4, c-Fos or FosB, inhibited this delayed stress response. EEF1D_L siRNA inhibited both the delayed and the extended primary stress responses, but had off target effects. In control cells an antioxidant response (ARE binding, HMOX1 mRNA levels) was detected 6 h after heat shock; in HSF1 K80Q cells this response was delayed to 24 h and the ARE complex had a different mobility. Inactivation of HSF1 thus affects the timing and nature of the antioxidant response and NRF2 can activate at least some HSF1 target genes in the absence of HSF1 activity.

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Abbreviations

HSP:

Heat shock protein

HSF1:

Heat shock factor 1

HSE:

Heat shock element

MEF:

Mouse embryonic fibroblast

NRF2:

Nuclear factor (erythroid-derived 2)-related factor 2

ARE:

Antioxidant response element

ATF:

Activating transcription factor

KEAP1:

Kelch-like ECH associated protein 1

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Acknowledgments

We thank Dr. A. Zantema for the HSPB1 antibody. We thank the microarray facility at the VU UMC (Amsterdam, The Netherlands) for performing the microarray experiments. This work was financially supported by AgentschapNL (project numbers IGE03018 and IGE07004, www.agentschapnl.nl).

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Authors and Affiliations

  1. 271 Department of Biomolecular Chemistry, Radboud University Nijmegen, P.O. Box 9101, 6500 HB, Nijmegen, The Netherlands

    Sanne M. M. Hensen, Lonneke Heldens, Siebe T. van Genesen, Ger J. M. Pruijn & Nicolette H. Lubsen

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  1. Sanne M. M. Hensen
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  2. Lonneke Heldens
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  3. Siebe T. van Genesen
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  4. Ger J. M. Pruijn
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  5. Nicolette H. Lubsen
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Correspondence to Nicolette H. Lubsen.

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Hensen, S.M.M., Heldens, L., van Genesen, S.T. et al. A delayed antioxidant response in heat-stressed cells expressing a non-DNA binding HSF1 mutant. Cell Stress and Chaperones 18, 455–473 (2013). https://doi.org/10.1007/s12192-012-0400-0

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  • DOI: https://doi.org/10.1007/s12192-012-0400-0

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