Functional diversification of closely related ARF-GEFs in protein secretion and recycling

Abstract

Guanine-nucleotide exchange factors on ADP-ribosylation factor GTPases (ARF-GEFs) regulate vesicle formation in time and space by activating ARF substrates on distinct donor membranes1. Mammalian GBF1 (ref. 2) and yeast Gea1/2 (ref. 3) ARF-GEFs act at Golgi membranes, regulating COPI-coated vesicle formation. In contrast, their Arabidopsis thaliana homologue GNOM (GN) is required for endosomal recycling, playing an important part in development4. This difference indicates an evolutionary divergence of trafficking pathways between animals and plants, and raised the question of how endoplasmic reticulum–Golgi transport is regulated in plants. Here we demonstrate that the closest homologue of GNOM in Arabidopsis, GNOM-LIKE1 (GNL1; NM_123312; At5g39500), performs this ancestral function. GNL1 localizes to and acts primarily at Golgi stacks, regulating COPI-coated vesicle formation. Surprisingly, GNOM can functionally substitute for GNL1, but not vice versa. Our results suggest that large ARF-GEFs of the GBF1 class perform a conserved role in endoplasmic reticulum–Golgi trafficking and secretion, which is done by GNL1 and GNOM in Arabidopsis, whereas GNOM has evolved to perform an additional plant-specific function of recycling from endosomes to the plasma membrane. Duplication and diversification of ARF-GEFs in plants contrasts with the evolution of entirely new classes of ARF-GEFs5 for endosomal trafficking in animals, which illustrates the independent evolution of complex endosomal pathways in the two kingdoms.

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Figure 1: Identification of the GNL1 compartment.
Figure 2: Localization of BFA-resistant and BFA-sensitive GNL1 after BFA treatment.
Figure 3: Phenotype of gnl1 mutants and relationship of GNL1 to GN.
Figure 4: Functional relationship between GN and GNL1.

References

  1. 1

    Shin, H. W. & Nakayama, K. Guanine nucleotide-exchange factors for arf GTPases: their diverse functions in membrane traffic. J. Biochem. 136, 761–767 (2004)

  2. 2

    Zhao, X. et al. GBF1, a cis-Golgi and VTCs-localized ARF-GEF, is implicated in ER-to-Golgi protein traffic. J. Cell Sci. 119, 3743–3753 (2006)

  3. 3

    Peyroche, A., Courbeyrette, R., Rambourg, A. & Jackson, C. L. The ARF exchange factors Gea1p and Gea2p regulate Golgi structure and function in yeast. J. Cell Sci. 114, 2241–2253 (2001)

  4. 4

    Geldner, N. et al. The Arabidopsis GNOM ARF-GEF mediates endosomal recycling, auxin transport, and auxin-dependent plant growth. Cell 112, 219–230 (2003)

  5. 5

    Cox, R., Mason-Gamer, R. J., Jackson, C. L. & Segev, N. Phylogenetic analysis of Sec7-domain-containing Arf nucleotide exchangers. Mol. Biol. Cell 15, 1487–1505 (2004)

  6. 6

    Mouratou, B. et al. The domain architecture of large guanine nucleotide exchange factors for the small GTP-binding protein Arf. BMC Genom. 6, 20 (2005)

  7. 7

    Spang, A., Herrmann, J. M., Hamamoto, S. & Schekman, R. The ADP ribosylation factor-nucleotide exchange factors Gea1p and Gea2p have overlapping, but not redundant functions in retrograde transport from the Golgi to the endoplasmic reticulum. Mol. Biol. Cell 12, 1035–1045 (2001)

  8. 8

    Mayer, U., Büttner, G. & Jürgens, G. Apical–basal pattern formation in the Arabidopsis embryo: studies on the role of the gnom gene. Development 117, 149–162 (1993)

  9. 9

    Steinmann, T. et al. Coordinated polar localization of auxin efflux carrier PIN1 by GNOM ARF GEF. Science 286, 316–318 (1999)

  10. 10

    Cherfils, J. & Melancon, P. On the action of Brefeldin A on Sec7-stimulated membrane-recruitment and GDP/GTP exchange of Arf proteins. Biochem. Soc. Trans. 33, 635–638 (2005)

  11. 11

    Peyroche, A. et al. Brefeldin A acts to stabilize an abortive ARF–GDP–Sec7 domain protein complex: involvement of specific residues of the Sec7 domain. Mol. Cell 3, 275–285 (1999)

  12. 12

    Barzilay, E., Ben-Califa, N., Hirschberg, K. & Neumann, D. Uncoupling of brefeldin A-mediated coatomer protein complex-I dissociation from Golgi redistribution. Traffic 6, 794–802 (2005)

  13. 13

    Niu, T. K., Pfeifer, A. C., Lippincott-Schwartz, J. & Jackson, C. L. Dynamics of GBF1, a Brefeldin A-sensitive Arf1 exchange factor at the Golgi. Mol. Biol. Cell 16, 1213–1222 (2005)

  14. 14

    Geldner, N., Friml, J., Stierhof, Y.-D., Jürgens, G. & Palme, K. Auxin transport inhibitors block PIN1 cycling and vesicle trafficking. Nature 413, 425–428 (2001)

  15. 15

    Schmid, M. et al. A gene expression map of Arabidopsis thaliana development. Nature Genet. 37, 501–506 (2005)

  16. 16

    Pimpl, P. et al. In situ localization and in vitro induction of plant COPI-coated vesicles. Plant Cell 12, 2219–2236 (2000)

  17. 17

    Dettmer, J., Hong-Hermesdorf, A., Stierhof, Y.-D. & Schumacher, K. Vacuolar H+-ATPase activity is required for endocytic and secretory trafficking in Arabidopsis. Plant Cell 18, 715–730 (2006)

  18. 18

    Ueda, T., Uemura, T., Sato, M. H. & Nakano, A. Functional differentiation of endosomes in Arabidopsis cells. Plant J. 40, 783–789 (2004)

  19. 19

    Lee, G. J., Sohn, E. J., Lee, M. H. & Hwang, I. The Arabidopsis rab5 homologs Rha1 and Ara7 localize to the prevacuolar compartment. Plant Cell Physiol. 45, 1211–1220 (2004)

  20. 20

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

  21. 21

    Teh, O. & Moore, I. An ARF-GEF acting at the Golgi and in selective endocytosis in polarized plant cells. Nature doi: 10.1038/nature06023 (this issue)

  22. 22

    Geldner, N. et al. Partial loss-of-function alleles reveal a role for GNOM in auxin transport-related, post-embryonic development of Arabidopsis. Development 131, 389–400 (2004)

  23. 23

    Ritzenthaler, C. et al. Reevaluation of the effects of brefeldin A on plant cells using tobacco Bright Yellow 2 cells expressing Golgi-targeted green fluorescent protein and COPI antisera. Plant Cell 14, 237–261 (2002)

  24. 24

    Shin, H. W., Morinaga, N., Noda, M. & Nakayama, K. BIG2, a guanine nucleotide exchange factor for ADP-ribosylation factors: its localization to recycling endosomes and implication in the endosome integrity. Mol. Biol. Cell 15, 5283–5294 (2004)

  25. 25

    Weijers, D., Van Hamburg, J. P., Van Rijn, E., Hooykaas, P. J. & Offringa, R. Diphtheria toxin-mediated cell ablation reveals interregional communication during Arabidopsis seed development. Plant Physiol. 133, 1882–1892 (2003)

  26. 26

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

  27. 27

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

  28. 28

    Völker, A., Stierhof, Y.-D. & Jürgens, G. Cell cycle-independent expression of the Arabidopsis cytokinesis-specific syntaxin KNOLLE results in mistargeting to the plasma membrane and is not sufficient for cytokinesis. J. Cell Sci. 114, 3001–3012 (2001)

  29. 29

    Rios, G. et al. Rapid identification of Arabidopsis insertion mutants by non-radioactive detection of T-DNA tagged genes. Plant J. 32, 243–253 (2002)

  30. 30

    Vieten, A. et al. Functional redundancy of PIN proteins is accompanied by auxin-dependent cross-regulation of PIN expression. Development 132, 4521–4531 (2005)

  31. 31

    Song, J., Lee, M. H., Lee, G. J., Yoo, C. M. & Hwang, I. Arabidopsis EPSIN1 plays an important role in vacuolar trafficking of soluble cargo proteins in plant cells via interactions with clathrin, AP-1, VTI11, and VSR1. Plant Cell 18, 2258–2274 (2006)

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Acknowledgements

We thank N. Takada and L. Müller for technical assistance, A. Vieten, J. Friml, F. El-Kasmi, G. Strompen and K. Steinborn for screening the Cologne T-DNA insertion lines, K. Schumacher, I. Hwang, M. Grebe, A. Schlereth and D.Weijers for providing materials, O. Teh and I. Moore for sharing unpublished material and results, and N. Anders, U. Mayer, K. Schumacher and D. Weigel for critically reading the manuscript and suggestions. We especially thank N. Anders for advice and discussions. This work was supported by an EMBO long-term Fellowship to J.S. and by grants from the Human Frontier in Science Program Organization and the SFB 446 of the Deutsche Forschungsgemeinschaft to G.J.

Author Contributions S.R. carried out most of the experiments, N.G. initiated the project, J.S. generated the RNAi and promoter-swap lines, H.W. generated the ARA7–GFP marker line, Y.-D.S. performed the electron microscopy analysis and immunogold localization experiments, G.R. and C.K. provided the T-DNA collection, D.G.R. generated antisera against markers, and G.J. and S.R. designed the experiments, discussed the results and wrote the manuscript.

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Correspondence to Gerd Jürgens.

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Richter, S., Geldner, N., Schrader, J. et al. Functional diversification of closely related ARF-GEFs in protein secretion and recycling. Nature 448, 488–492 (2007). https://doi.org/10.1038/nature05967

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