Letter | Published:

Sterol-dependent endocytosis mediates post-cytokinetic acquisition of PIN2 auxin efflux carrier polarity

Nature Cell Biology volume 10, pages 237244 (2008) | Download Citation

Subjects

Abstract

The polarization of yeast and animal cells relies on membrane sterols for polar targeting of proteins to the plasma membrane, their polar endocytic recycling and restricted lateral diffusion1,2,3,4. However, little is known about sterol function in plant-cell polarity5. Directional root growth along the gravity vector requires polar transport of the plant hormone auxin. In Arabidopsis, asymmetric plasma membrane localization of the PIN–FORMED2 (PIN2) auxin transporter directs root gravitropism6,7,8,9,10. Although the composition of membrane sterols influences gravitropism and localization of two other PIN proteins11, it remains unknown how sterols contribute mechanistically to PIN polarity. Here, we show that correct membrane sterol composition is essential for the acquisition of PIN2 polarity. Polar PIN2 localization is defective in the sterol-biosynthesis mutant cyclopropylsterol isomerase1-1 (cpi1-1) which displays altered sterol composition, PIN2 endocytosis, and root gravitropism. At the end of cytokinesis, PIN2 localizes initially to both newly formed membranes but subsequently disappears from one. By contrast, PIN2 frequently remains at both daughter membranes in endocytosis-defective cpi1-1 cells. Hence, sterol composition affects post-cytokinetic acquisition of PIN2 polarity by endocytosis, suggesting a mechanism for sterol action on establishment of asymmetric protein localization.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Accessions

GenBank/EMBL/DDBJ

References

  1. 1.

    & Cholesterol is required for surface transport of influenza virus hemagglutinin. J. Cell Biol. 140, 1357–1367 (1998).

  2. 2.

    & Cell surface polarization during yeast mating. Proc. Natl Acad. Sci. USA 99, 14183–14188 (2002).

  3. 3.

    & Slow diffusion of proteins in the yeast plasma membrane allows polarity to be maintained by endocytic cycling. Curr. Biol. 13, 1636–1640 (2003).

  4. 4.

    & Where sterols are required for endocytosis. Biochim. Biophys. Acta 1666, 51–61 (2004).

  5. 5.

    & Plant cell polarity: the ins-and-outs of sterol transport. Curr. Biol. 13, 781–783 (2003).

  6. 6.

    , , & EIR1, a root-specific protein involved in auxin transport, is required for gravitropism in Arabidopsis thaliana. Genes Dev. 12, 2175–2187 (1998).

  7. 7.

    , , & AGR, an agravitropic locus of Arabidopsis thaliana, encodes a novel membrane-protein family member. Plant Cell Physiol. 39, 1111–1118 (1998).

  8. 8.

    et al. The Arabidopsis thaliana AGRAVITROPIC 1 gene encodes a component of the polar-auxin-transport efflux carrier. Proc. Natl Acad. Sci. USA 95, 15112–15117 (1998).

  9. 9.

    et al. AtPIN2 defines a locus of Arabidopsis for root gravitropism control. EMBO J. 17, 6903–6911 (1998).

  10. 10.

    et al. Polar PIN localization directs auxin flow in plants. Science 312, 883 (2006).

  11. 11.

    et al. Cell polarity and PIN protein positioning in Arabidopsis require STEROL METHYLTRANSFERASE1 function. Plant Cell 15, 612–625 (2003).

  12. 12.

    et al. PIN proteins perform a rate-limiting function in cellular auxin efflux. Science 312, 914–918 (2006).

  13. 13.

    , , & Molecular and cellular aspects of auxin-transport-mediated development. Trends Plant Sci. 12, 160–168 (2007).

  14. 14.

    et al. hydra mutants of Arabidopsis are defective in sterol profiles and auxin and ethylene signaling. Plant Cell 14, 1017–1031 (2002).

  15. 15.

    et al. Arabidopsis sterol endocytosis involves actin-mediated trafficking via ARA6-positive early endosomes. Curr. Biol. 13, 1378–1387 (2003).

  16. 16.

    , , , & Functional cloning of an Arabidopsis thaliana cDNA encoding cycloeucalenol cycloisomerase. J. Biol. Chem. 275, 13394–13397 (2000).

  17. 17.

    et al. Root gravitropism requires lateral root cap and epidermal cells for transport and response to a mobile auxin signal. Nature Cell Biol. 7, 1057–1065 (2005).

  18. 18.

    et al. An auxin-dependent distal organizer of pattern and polarity in the Arabidopsis root. Cell 99, 463–472 (1999).

  19. 19.

    et al. Vectorial information for Arabidopsis planar polarity is mediated by combined AUX1, EIN2, and GNOM activity. Curr. Biol. 16, 2143–2149 (2006).

  20. 20.

    et al. The Arabidopsis KNOLLE protein is a cytokinesis-specific syntaxin. J. Cell Biol. 139, 1485–1493 (1997).

  21. 21.

    & Plant membrane sterols: isolation, identification, and biosynthesis. Methods Enzymol. 148, 632–650 (1987).

  22. 22.

    & Dissection of Arabidopsis ADP-RIBOSYLATION FACTOR 1 function in epidermal cell polarity. Plant Cell 17, 525–536 (2005).

  23. 23.

    et al. Syntaxin specificity of cytokinesis in Arabidopsis. Nature Cell Biol. 5, 531–534 (2003).

  24. 24.

    , , & Ara6, a plant-unique novel type Rab GTPase, functions in the endocytic pathway of Arabidopsis thaliana. EMBO J. 20, 4730–4741 (2001).

  25. 25.

    et al. Clathrin-mediated constitutive endocytosis of PIN auxin efflux carriers in Arabidopsis. Curr. Biol. 17, 520–527 (2007).

  26. 26.

    , , & Filipin-sensitive caveolae-mediated transport in endothelium: reduced transcytosis, scavenger endocytosis, and capillary permeability of select macromolecules. J. Cell Biol. 127, 1217–1232 (1994).

  27. 27.

    , , , & Subcellular trafficking of the Arabidopsis auxin influx carrier AUX1 uses a novel pathway distinct from PIN1. Plant Cell 18, 3171–3181 (2006).

  28. 28.

    et al. Lipid rafts in higher plant cells: purification and characterization of Triton X-100-insoluble microdomains from tobacco plasma membrane. J. Biol. Chem. 279, 36277–36286 (2004).

  29. 29.

    et al. Analysis of detergent-resistant membranes in Arabidopsis. Evidence for plasma membrane lipid rafts. Plant Physiol. 137, 104–116 (2005).

  30. 30.

    et al. Insights into the role of specific lipids in the formation and delivery of lipid microdomains to the plasma membrane of plant cells. Plant Physiol. 143, 461–472 (2007).

Download references

Acknowledgements

We gratefully acknowledge M. Bennett, D. Ehrhardt, T. Guilfoyle, J. Haseloff, R. Heidstra, R. Hellens, I. Moore, P. Mullineaux, G. Jürgens, B. Scheres, R. Swarup and J. Xu for sharing published research materials used in this study. We also acknowledge the Nottingham Arabidopsis Stock Centre for distributing mutant lines including SALK T-DNA insertion mutants, provided by J. Alonso and J. Ecker, and the John Innes Centre for EXOTIC Gene Trap lines. We thank I.-B. Carlsson and K. Lundgren for technical assistance with sterol measurements and K. Schumacher for providing seedlings for electron microscopy. We thank L. Bako, R. Bhalerao, U. Fischer, E. Johnson, A. Marchant, and G. Samuelsson for discussions and comments on the manuscript. This work was supported by a grant from the Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning (Formas) to M.G., in part by a postdoctoral stipend from the Carl Tryggers Foundation to Y.B., an EU Marie-Curie International Incoming Postdoctoral Fellowship to Y.I., and the Swedish Foundation for Strategic Research (SSF).

Author information

Author notes

    • Markus Grebe

    Present address: Umeå Plant Science Centre, Department of Plant Physiology, University of Umeå, SE-90187 Umeå, Sweden.

Affiliations

  1. Umeå Plant Science Centre (UPSC), Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, SE-90183 Umeå, Sweden.

    • Shuzhen Men
    • , Yohann Boutté
    • , Yoshihisa Ikeda
    • , Thomas Moritz
    •  & Markus Grebe
  2. Institute of Biology II, University of Freiburg, Schänzlestrasse 1, D-79104 Freiburg, Germany.

    • Xugang Li
    •  & Klaus Palme
  3. Centre for Plant Molecular Biology, University of Tübingen, Auf der Morgenstelle 5, D-72076 Tübingen, Germany.

    • York-Dieter Stierhof
  4. Institute for Plant Molecular Biology (IBMP, UPR CNRS 2357), Université Louis Pasteur, 28 rue Goethe, F-67083 Strasbourg, France.

    • Marie-Andrée Hartmann

Authors

  1. Search for Shuzhen Men in:

  2. Search for Yohann Boutté in:

  3. Search for Yoshihisa Ikeda in:

  4. Search for Xugang Li in:

  5. Search for Klaus Palme in:

  6. Search for York-Dieter Stierhof in:

  7. Search for Marie-Andrée Hartmann in:

  8. Search for Thomas Moritz in:

  9. Search for Markus Grebe in:

Corresponding author

Correspondence to Markus Grebe.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary figures S1, S2, S3, S4, S5, Supplementary table S1 and Supplementary Methods

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ncb1686

Further reading