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Caffeine inhibits PI3K and mTORC2 in Dictyostelium and differentially affects multiple other cAMP chemoattractant signaling effectors

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Abstract

Caffeine is commonly used in Dictyostelium to inhibit the synthesis of the chemoattractant cAMP and, therefore, its secretion and the autocrine stimulation of cells, in order to prevent its interference with the study of chemoattractant-induced responses. However, the mechanism through which caffeine inhibits cAMP synthesis in Dictyostelium has not been characterized. Here, we report the effects of caffeine on the cAMP chemoattractant signaling network. We found that caffeine inhibits phosphatidylinositol 3-kinase (PI3K) and mechanistic target of rapamycin complex 2 (mTORC2). Both PI3K and mTORC2 are essential for the chemoattractant-stimulated cAMP production, thereby providing a mechanism for the caffeine-mediated inhibition of cAMP synthesis. Our results also reveal that caffeine treatment of cells leads to an increase in cAMP-induced RasG and Rap1 activation, and inhibition of the PKA, cGMP, MyoII, and ERK1 responses. Finally, we observed that caffeine has opposite effects on F-actin and ERK2 depending on the assay and Dictyostelium strain used, respectively. Altogether, our findings reveal that caffeine considerably affects the cAMP-induced chemotactic signaling pathways in Dictyostelium, most likely acting through multiple targets that include PI3K and mTORC2.

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Acknowledgements

We are grateful to the Dicty Stock Center and its material depositors for providing cells and DNA constructs.

Funding

This study was funded by a Research Scholar Grant 127940-RSG-15-024-01-CSM from the American Cancer Society to P.G.C. M.S. was supported by NIH T32 Grant GM008804 and P.L. was supported by a U.S. Public Health Service Grant GM037830.

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Author notes

  1. Margarethakay Scavello

    Present address: Eurofins Lancaster Laboratories Professional Scientific Services, LLC, Malvern, PA, USA

  2. Dustin Daniel

    Present address: Department of Basic Medical Sciences, University of Arizona, Phoenix, AZ, USA

  3. Pearce Haldeman

    Present address: Division of Biology and Biological Engineering, Joint Center for Transitional Medicine, California Institute of Technology, Pasadena, CA, USA

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Arizona, Tucson, AZ, USA

    A. F. M. Tariqul Islam, Margarethakay Scavello, Pouya Lotfi, Dustin Daniel, Pearce Haldeman & Pascale G. Charest

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Correspondence to Pascale G. Charest.

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Tariqul Islam, A.F.M., Scavello, M., Lotfi, P. et al. Caffeine inhibits PI3K and mTORC2 in Dictyostelium and differentially affects multiple other cAMP chemoattractant signaling effectors. Mol Cell Biochem 457, 157–168 (2019). https://doi.org/10.1007/s11010-019-03520-z

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