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Phosphoinositide phosphatase activity coupled to an intrinsic voltage sensor

Nature volume 435, pages 1239–1243 (2005)Cite this article

Abstract

Changes in membrane potential affect ion channels and transporters, which then alter intracellular chemical conditions. Other signalling pathways coupled to membrane potential have been suggested1,2,3 but their underlying mechanisms are unknown. Here we describe a novel protein from the ascidian Ciona intestinalis that has a transmembrane voltage-sensing domain homologous to the S1–S4 segments of voltage-gated channels and a cytoplasmic domain similar to phosphatase and tensin homologue. This protein, named C. intestinalis voltage-sensor-containing phosphatase (Ci-VSP), displays channel-like ‘gating’ currents and directly translates changes in membrane potential into the turnover of phosphoinositides. The activity of the phosphoinositide phosphatase in Ci-VSP is tuned within a physiological range of membrane potential. Immunocytochemical studies show that Ci-VSP is expressed in Ciona sperm tail membranes, indicating a possible role in sperm function or morphology. Our data demonstrate that voltage sensing can function beyond channel proteins and thus more ubiquitously than previously realized.

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Figure 1: Transmembrane domain of Ci-VSP operates as a voltage sensor.
Figure 2: Cytoplasmic domain of Ci-VSP is a phosphoinositide phosphatase.
Figure 3: Ci-VSP alters phosphoinositide concentration in a voltage-dependent manner as probed with K + channel activities.
Figure 4: Ci-VSP protein is expressed in sperm tail.

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Acknowledgements

We thank D. E. Logothetis for providing IRK1 R228Q plasmid, Y. Kubo for providing wild-type IRK1 plasmid, T. Nukada for G-protein β1 and γ1 subunit plasmids, D. McKinnon and K. Nakajo for KCNQ2/3 plasmids, M. Lazdunski for GIRK2 plasmid, T. Maehama for advice on measuring phosphatase activity, F. Kukita for help in cut-open oocyte recording, J. Cui for discussion, N. Satoh and Y. Satou for help in bioinformatics, and D. McLean for critical reading of the manuscript. This work was supported by Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists to Y.M., Grants-in-Aid for Scientific Research to Y.O. and H.I., and a Grant-in-Aid for Creative Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology to Y.O.

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Author notes

  1. Yoshimichi Murata, Hirohide Iwasaki and Mari Sasaki: *These authors contributed equally to this work

Authors and Affiliations

  1. Section of Developmental Neurophysiology, Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Higashiyama 5-1, Myodaiji-cho, 444-8787, Aichi, Okazaki, Japan

    Yoshimichi Murata, Hirohide Iwasaki, Mari Sasaki & Yasushi Okamura

  2. National Institute for Physiological Sciences, National Institutes of Natural Sciences, Nishigonaka 38, Myodaiji-cho, 444-8585, Aichi, Okazaki, Japan

    Yoshimichi Murata, Hirohide Iwasaki, Mari Sasaki & Yasushi Okamura

  3. School of Life Science, The Graduate University for Advanced Studies, Okazaki, 444-8585, Japan

    Hirohide Iwasaki, Mari Sasaki & Yasushi Okamura

  4. Shimoda Marine Research Center, Graduate School of Life and Environmental Sciences, University of Tsukuba, 5-10-1 Shimoda, 415-0025, Shizuoka, Japan

    Kazuo Inaba

  5. Neuroscience Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba Central 6, Higashi 1-1-1, 305-8566, Ibaraki, Tsukuba, Japan

    Yasushi Okamura

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  1. Yoshimichi Murata
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Corresponding author

Correspondence to Yasushi Okamura.

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Competing interests

The full-length cDNA sequence of Ci-VSP is deposited in the DNA Data Bank of Japan (DDBJ) under the accession number AB183035. Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Figure S1

This figure illustrates coupling of voltage sensor and phosphatase revealed by mutated IRK1 channel and KCNQ2/3 channel. (DOC 180 kb)

Supplementary Figure S2

This illustrates effect of deletion of a linker region between voltage sensor and phosphatase domain of Ci-VSP. (DOC 142 kb)

Supplementary Figure S3

This illustrates specificity of antibodies for Ci-VSP. (DOC 231 kb)

Supplementary Method

This describes cloning of Ci-VSP cDNA, mutagenesis of Ci-VSP cDNA, Q-V plot of asymmetrical charge movements, biotinylation labeling of Ci-VSP protein in Xenopus oocyte, imunoblot analysis, in vitro phosphatase assay, RT-PCR detection of Ci-VSP transcript in Ciona intestinalis, and immunoelectron microscopy. (DOC 59 kb)

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Murata, Y., Iwasaki, H., Sasaki, M. et al. Phosphoinositide phosphatase activity coupled to an intrinsic voltage sensor. Nature 435, 1239–1243 (2005). https://doi.org/10.1038/nature03650

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