Phospholipase Cβ1 induces membrane tubulation and is involved in caveolae formation

Takehiko Inaba, Takuma Kishimoto, Motohide Murate, Takuya Tajima, Shota Sakai, Mitsuhiro Abe, Asami Makino, Nario Tomishige, Reiko Ishitsuka, Yasuo Ikeda, Shinji Takeoka, Toshihide Kobayashi

    Research output: Contribution to journalArticle

    14 Citations (Scopus)

    Abstract

    Lipid membrane curvature plays important roles in various physiological phenomena. Curvature-regulated dynamic membrane remodeling is achieved by the interaction between lipids and proteins. So far, several membrane sensing/sculpting proteins, such as Bin/amphiphysin/Rvs (BAR) proteins, are reported, but there remains the possibility of the existence of unidentified membrane-deforming proteins that have not been uncovered by sequence homology. To identify new lipid membrane deformation proteins, we applied liposome-based microscopic screening, using unbiased-darkfield microscopy. Using this method, we identified phospholipase Cβ1 (PLCβ1) as a new candidate. PLCβ1 is well characterized as an enzyme catalyzing the hydrolysis of phosphatidylinositol-4,5-bisphosphate (PIP2). In addition to lipase activity, our results indicate that PLCβ1 possessed the ability of membrane tubulation. Lipase domains and inositol phospholipids binding the pleckstrin homology (PH) domain of PLCβ1 were not involved, but the C-terminal sequence was responsible for this tubulation activity. Computational modeling revealed that the C terminus displays the structural homology to the BAR domains, which is well known as a membrane sensing/sculpting domain. Overexpression of PLCβ1 caused plasma membrane tubulation, whereas knockdown of the protein reduced the number of caveolae and induced the evagination of caveolin-rich membrane domains. Taken together, our results suggest a new function of PLCβ1: plasma membrane remodeling, and in particular, caveolae formation.

    Original languageEnglish
    Pages (from-to)7834-7839
    Number of pages6
    JournalProceedings of the National Academy of Sciences of the United States of America
    Volume113
    Issue number28
    DOIs
    Publication statusPublished - 2016 Jul 12

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    Keywords

    • BAR-like domain
    • Caveolae
    • Membrane tubulation
    • Microscopy screening
    • Phospholipase Cβ1

    ASJC Scopus subject areas

    • General

    Cite this

    Inaba, T., Kishimoto, T., Murate, M., Tajima, T., Sakai, S., Abe, M., Makino, A., Tomishige, N., Ishitsuka, R., Ikeda, Y., Takeoka, S., & Kobayashi, T. (2016). Phospholipase Cβ1 induces membrane tubulation and is involved in caveolae formation. Proceedings of the National Academy of Sciences of the United States of America, 113(28), 7834-7839. https://doi.org/10.1073/pnas.1603513113