Continuous light exposure causes cumulative stress that affects the localization oscillation dynamics of the transcription factor Msn2p.
Journal article, 2011

Light exposure is a potentially powerful stress factor during in vivo optical microscopy studies. In yeast, the general transcription factor Msn2p translocates from the cytoplasm to the nucleus in response to illumination. However, previous time-lapse fluorescence microscopy studies of Msn2p have utilized a variety of discrete exposure settings, which makes it difficult to correlate stress levels and illumination parameters. We here investigate how continuous illumination with blue light, corresponding to GFP excitation wavelengths, affects the localization pattern of Msn2p-GFP in budding yeast. The localization pattern was analyzed using a novel approach that combines wavelet decomposition and change point analysis. It was found that the Msn2p nucleocytoplasmic localization trajectories for individual cells exhibit up to three distinct and successive states; i) Msn2p localizes to the cytoplasm; ii) Msn2p rapidly shuttles between the cytoplasm and the nucleus; iii) Msn2p localizes to the nucleus. Many cells pass through all states consecutively at high light intensities, while at lower light intensities most cells only reach states i) or ii). This behaviour strongly indicates that continuous light exposure gradually increases the stress level over time, presumably through continuous accumulation of toxic photoproducts, thereby forcing the cell through a bistable region corresponding to nucleocytoplasmic oscillations. We also show that the localization patterns are dependent on protein kinase A (PKA) activity, i.e. yeast cells with constantly low PKA activity showed a stronger stress response. In particular, the nucleocytoplasmic oscillation frequency was found to be significantly higher for cells with low PKA activity for all light intensities.


Kristofer Bodvard

Chalmers, Applied Physics, Bionanophotonics

David Wrangborg

Fraunhofer-Chalmers Centre

Sofia Tapani

University of Gothenburg

Chalmers, Mathematical Sciences, Mathematical Statistics

Katarina Logg

University of Gothenburg

Chalmers, Applied Physics, Bionanophotonics

Piotr Sliwa

University of Gothenburg

Anders Blomberg

University of Gothenburg

Mats Kvarnström

Fraunhofer-Chalmers Centre

Mikael Käll

Chalmers, Applied Physics, Bionanophotonics

Biochimica et biophysica acta

0006-3002 (ISSN)

Vol. 1813 2 358-366

Subject Categories

Biochemistry and Molecular Biology





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