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Ionic leakage underlies a gain-of-function effect of dominant disease mutations affecting diverse P-type ATPases

Nature Genetics volume 46, pages 144–151 (2014)Cite this article

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Abstract

Type II P-type ATPases (PAIIs) constitute a family of conserved proteins that actively generate ionic gradients across membranes. Mutations in genes encoding PAIIs can cause heritable dominant diseases, with suggested etiology of haploinsufficiency. Using a Drosophila melanogaster genetic screen, we identified a dominant mutation altering the PAII member sarcoendoplasmic reticulum Ca2+ ATPase (SERCA). This mutation conferred temperature-sensitive uncoordination in a gain-of-function manner. We established that this gain-of-function phenotype is linked to dominant disease-causing mutations affecting various human PAIIs. We further found that heterologous expression of mutant PAIIs elicited ion leakage that was exacerbated at elevated temperatures. Therefore, these dominant mutations result in ionic leakage and render PAIIs susceptible to deleterious effects from elevated temperatures. Accordingly, it was recently reported that missense mutations affecting the Na+/K+ ATPase can elicit ionic leakage. We propose that ionic leakage is a pervasive gain-of-function mechanism that can underlie a variety of dominant PAII-related diseases.

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Figure 1: Amino acids altered in dominant PAII-related diseases in humans and alteration in Uncts2 mutant Drosophila.
Figure 2: Effects of dSERCA dominant alterations and elevated temperature on Ca2+ levels in intracellular stores.
Figure 3: Effects of mSERCA2 dominant alterations and elevated temperature on Ca2+ levels in intracellular stores.
Figure 4: Disease-causing alterations in the N and A domains elicit ionic leakage.
Figure 5: Dominant alterations in rPMCA2 elicit ionic leakage.
Figure 6: Effects of PMCA inhibition by increasing the pH to 9.0.

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Acknowledgements

We thank P. Gillespie (Oregon Health and Science University), T. Avidor-Reiss (University of Toledo), A. Kiger (University of California, San Diego), S. Courtneidge (Sanford-Burnham Medical Research Institute) and L. Luo (Stanford University) for reagents used in this work. We thank K. Spencer for technical assistance and A. Chadha for comments on the manuscript. The Uncts2 mutant was identified by screening a collection of F3 Drosophila lines in the laboratory of C. Zuker.

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  1. Department of Molecular and Cellular Neuroscience, Scripps Research Institute, La Jolla, California, USA

    Maki Kaneko, Bela S Desai & Boaz Cook

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  1. Maki Kaneko
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Contributions

B.C. performed mutagenesis screening and mapped the revertant lines. B.S.D. cloned and sequenced the dSERCA gene from the Uncts2 mutant. M.K. generated and tested all transgenic lines, sequenced dSERCA from additional temperature-sensitive uncoordinated mutants and performed the calcium-imaging experiments in cultured cells. B.C. led the entire project. M.K. generated tables and figures. M.K. and B.C. wrote the manuscript. All authors participated in the discussion and interpretation of data and results, and all participated in the editing and revision of the manuscript.

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Correspondence to Boaz Cook.

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Kaneko, M., Desai, B. & Cook, B. Ionic leakage underlies a gain-of-function effect of dominant disease mutations affecting diverse P-type ATPases. Nat Genet 46, 144–151 (2014). https://doi.org/10.1038/ng.2850

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