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Membrane stress is coupled to a rapid translational control of gene expression in chlorpromazine-treated cells

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Abstract

Chlorpromazine (CPZ) is a small permeable cationic amphiphilic molecule that inserts into membrane bilayers and binds to anionic lipids such as poly-phosphoinositides (PIs). Since PIs play important roles in many cellular processes, including signaling and membrane trafficking pathways, it has been proposed that CPZ affects cellular growth functions by preventing the recruitment of proteins with specific PI-binding domains. In this study, we have investigated the biological effects of CPZ in the yeast Saccharomyces cerevisiae. We screened a collection of approximately 4,800 gene knockout mutants, and found that mutants defective in membrane trafficking between the late-Golgi and endosomal compartments are highly sensitive to CPZ. Microscopy and transport analyses revealed that CPZ affects membrane structure of organelles, blocks membrane transport and activates the unfolded protein response (UPR). In addition, CPZ-treatment induces phosphorylation of the translation initiation factor (eIF2α), which reduces the general rate of protein synthesis and stimulates the production of Gcn4p, a major transcription factor that is activated in response to environmental stresses. Altogether, our results reveal that membrane stress within the cells rapidly activates an important gene expression program, which is followed by a general inhibition of protein synthesis. Remarkably, the increase of phosphorylated eIF2α and protein synthesis inhibition were also detected in CPZ-treated NIH-3T3 fibroblasts, suggesting the existence of a conserved mechanism of translational regulation that operates during a membrane stress.

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Abbreviations

CPZ :

Chlorpromazine

Tm :

Tunicamycin

3-AT :

3-Aminotriazole

UPR :

Unfolded protein response

ER :

Endoplasmic reticulum

PI :

Phosphoinositide

PS :

Phosphatidylserine

PA :

Phosphatidylamine

PE :

Phosphatidylethanolamine

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Acknowledgments

We are extremely grateful for continuous support from Costa Georgopoulos. We thank S. Emr, T. R. Graham, R. Schekman, A. Hinnebusch and T. Dever for providing strains, plasmids and antibodies. We also thank D. Ang for a critical reading of the manuscript and the PFMU at the Geneva Medical Faculty for access to electron microscope and ancillary equipments. This work was supported by grants from the Swiss National Science Foundation and the Canton of Geneva to C. Georgopoulos (FN-31-654039), M.F. (FN-3100A0-104489), PL (FN-3100A0-105894/1) and CDV (PP00A-106754/1).

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  1. Elisabetta Cameroni & Claudio De Virgilio

    Present address: Département de Médecine, Division de Biochimie, Université de Fribourg, 1700, Fribourg, Switzerland

Authors and Affiliations

  1. Département de Microbiologie et Médecine Moléculaire, Centre Médical Universitaire, Université de Genève, 1, rue Michel-Servet, 1211, Geneva 4, Switzerland

    Loic De Filippi, Elisabetta Cameroni, Patrick Linder, Claudio De Virgilio & Olivier Deloche

  2. Département de Physiologie Cellulaire et Métabolisme, CMU, Université de Genève, 1211, Geneva 4, Switzerland

    Margot Fournier & Michelangelo Foti

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  1. Loic De Filippi
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  2. Margot Fournier
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  3. Elisabetta Cameroni
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Correspondence to Olivier Deloche.

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Communicated by Gerhard Braus.

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De Filippi, L., Fournier, M., Cameroni, E. et al. Membrane stress is coupled to a rapid translational control of gene expression in chlorpromazine-treated cells. Curr Genet 52, 171–185 (2007). https://doi.org/10.1007/s00294-007-0151-0

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