Cellular response to heat shock studied by multiconfocal fluorescence correlation spectroscopy.

Heat shock triggers a transient and ubiquitous response, the function of which is to protect cells against stress-induced damage. The heat-shock response is controlled by a key transcription factor known as heat shock factor 1 (HSF1). We have developed a multiconfocal fluorescence correlation spectroscopy setup to measure the dynamics of HSF1 during the course of the heat-shock response. The system combines a spatial light modulator, to address several points of interest, and an electron-multiplying charge-coupled camera for fast multiconfocal recording of the photon streams. Autocorrelation curves with a temporal resolution of 14 μs were analyzed before and after heat shock on eGFP and HSF1-eGFP-expressing cells. Evaluation of the dynamic parameters of a diffusion-and-binding model showed a slower HSF1 diffusion after heat shock. It is also observed that the dissociation rate decreases after heat shock, whereas the association rate is not affected. In addition, thanks to the multiconfocal fluorescence correlation spectroscopy system, up to five spots could be simultaneously located in each cell nucleus. This made it possible to quantify the intracellular variability of the diffusion constant of HSF1, which is higher than that of inert eGFP molecules and increases after heat shock. This finding is consistent with the fact that heat-shock response is associated with an increase of HSF1 interactions with DNA and cannot be explained even partially by heat-induced modifications of nuclear organization.

Domaines

Biophysique [physics.bio-ph] Biologie cellulaire

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mFCS-manuscript-Delon.pdf (9.56 Mo)

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mFCS-Supp-mat-Delon.pdf (491.61 Ko)

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https://hal.science/hal-00849356

Soumis le : mardi 30 juillet 2013-17:48:50

Dernière modification le : jeudi 4 avril 2024-21:28:49

Archivage à long terme le : mercredi 5 avril 2017-18:19:07

Dates et versions

hal-00849356 , version 1 (30-07-2013)

Identifiants

HAL Id : hal-00849356 , version 1
DOI : 10.1016/j.bpj.2012.07.041
PUBMED : 22995483

Citer

Meike Kloster-Landsberg, Gaëtan Herbomel, Irène Wang, Jacques Derouard, Claire Vourc'H, et al.. Cellular response to heat shock studied by multiconfocal fluorescence correlation spectroscopy.. Biophysical Journal, 2012, 103 (6), pp.1110-9. ⟨10.1016/j.bpj.2012.07.041⟩. ⟨hal-00849356⟩

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