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GPHR is a novel anion channel critical for acidification and functions of the Golgi apparatus

Nature Cell Biology volume 10, pages 1135–1145 (2008)Cite this article

Abstract

The organelles within secretory and endocytotic pathways in mammalian cells have acidified lumens, and regulation of their acidic pH is critical for the trafficking, processing and glycosylation of cargo proteins and lipids, as well as the morphological integrity of the organelles. How organelle lumen acidification is regulated, and how luminal pH elevation disturbs these fundamental cellular processes, is largely unknown. Here, we describe a novel molecule involved in Golgi acidification. First, mutant cells defective in Golgi acidification were established that exhibited delayed protein transport, impaired glycosylation and Golgi disorganization. Using expression cloning, a novel Golgi-resident multi-transmembrane protein, named Golgi pH regulator (GPHR), was identified as being responsible for the mutant cells. After reconstitution in planar lipid bilayers, GPHR exhibited a voltage-dependent anion-channel activity that may function in counterion conductance. Thus, GPHR modulates Golgi functions through regulation of acidification.

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Figure 1: Delayed transport of newly synthesized proteins in mutant cells.
Figure 2: Impaired glycosylation and Golgi disorganization in mutant cells.
Figure 3: GPHR is responsible for the defective phenotypes in C27 cells.
Figure 4: Impaired luminal acidification of the Golgi is the primary defect in C27 cells.
Figure 5: GPHR is mainly localized at the Golgi.
Figure 6: GPHR is an anion channel.
Figure 7: Trimeric GPHR channel helps V-ATPase to acidify the Golgi lumen.

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Acknowledgements

We thank F. Mori and K. Kinoshita for excellent technical assistance, and K. Nakamura for help with the cell sorting. We also thank T. Yoshimori, S. Kimura and H. Oomori for allowing us to use confocal and electron microscopes and for technical direction, and H. Hibino, H. Takeshima and T. Yamazaki for helpful discussions. This work was supported by grants from the Precursory Research for Embryonic Science and Technology, Japan Science and Technology Agency (Y.M.), the Core Research for Evolutional Science and Technology, Japan Science and Technology Agency (T.K.) and the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Author notes

  1. Masato Koike & Yasuo Uchiyama

    Present address: Current address: Department of Cell Biology and Neuroscience, Juntendo University School of Medicine, Tokyo 113–8421, Japan.,

Authors and Affiliations

  1. Research Institute for Microbial Diseases, Osaka University, Suita, 565–0871, Osaka, Japan

    Yusuke Maeda & Taroh Kinoshita

  2. WPI Immunology Frontier Research Center, Osaka University, Suita, 565–0871, Osaka, Japan

    Yusuke Maeda & Taroh Kinoshita

  3. PRESTO Japan Science and Technology Agency, Saitama, 332–0012, Japan

    Yusuke Maeda

  4. Laboratories for Nanobiology, Graduate School of Frontier Biosciences, Osaka University, Suita, 565–0871, Osaka, Japan

    Toru Ide

  5. Molecular-Informational Life Science Research Group, RIKEN, 351–0198, Wako, Japan

    Toru Ide

  6. Department of Cell Biology and Neurosciences, Graduate School of Medicine, Osaka University, Suita, 565–0871, Osaka, Japan

    Masato Koike & Yasuo Uchiyama

  7. CREST, Japan Science and Technology Agency, Saitama, 332–0012, Japan

    Taroh Kinoshita

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Contributions

T.I. performed the planar lipid-bilayer analyses. M.K. and Y.U. performed the electron microscopy analyses. Y.M. performed the other experiments. T.K. and Y.M. contributed to the planning of experiments and wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Yusuke Maeda.

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Supplementary Figures S1, S2, S3, S4, S5, S6, S7, S8, Supplementary Materials and Methods (PDF 2164 kb)

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Maeda, Y., Ide, T., Koike, M. et al. GPHR is a novel anion channel critical for acidification and functions of the Golgi apparatus. Nat Cell Biol 10, 1135–1145 (2008). https://doi.org/10.1038/ncb1773

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