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TRAPPII subunits are required for the specificity switch of a Ypt–Rab GEF

Nature Cell Biology volume 8, pages 1263–1269 (2006)Cite this article

Abstract

Ypt–Rab GTPases are key regulators of the various steps of intracellular trafficking. Guanine nucleotide-exchange factors (GEFs) regulate the conversion of Ypt–Rabs to the GTP-bound state, in which they interact with effectors that mediate all the known aspects of vesicular transport1,2,3. An interesting possibility is that Ypt–Rabs coordinate separate steps of the transport pathways4. The conserved modular complex TRAPP is a GEF for the Golgi gatekeepers Ypt1 and Ypt31/32 (Refs 5–7). However, it is not known how Golgi entry and exit are coordinated. TRAPP comes in two configurations: the seven-subunit TRAPPI is required for endoplasmic reticulum-to-Golgi transport, whereas the ten-subunit TRAPPII functions in late Golgi8,9,10. The two essential TRAPPII-specific subunits Trs120 and Trs130 have been identified as Ypt31/32 genetic interactors11,12,13. Here, we show that they are required for switching the GEF specificity of TRAPP from Ypt1 to Ypt31. Moreover, a trs130ts mutation confers opposite effects on the intracellular localization of these GTPases. We suggest that the Trs120–Trs130 subcomplex joins TRAPP in the late Golgi to switch its GEF activity from Ypt1 to Ypt31/32. Such a 'switchable' GEF could ensure sequential activation of these Ypts, thereby coordinating Golgi entry and exit.

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Figure 1: Purification of GST–Bet5-associated complexes from wild-type (WT) and trs130- and trs120-mutant cells.
Figure 2: TRAPP purified from trs130- and trs120-mutant cells is defective in Ypt31 GEF, but possesses higher Ypt1 GEF activity than wild-type TRAPP.
Figure 3: TRAPPII-specific subunits regulate Ypt1 and Ypt31 in opposite ways in vitro and in vivo.
Figure 4: Physical and genetic interactions between TRAPP subunits and the Golgi Ypts.
Figure 5: Models for the specificity-switch of the dual TRAPP GEF by Trs130.

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Acknowledgements

We thank D. Stone for critical reading of the manuscript, A. Lodhi for tehcnical help and Y. Jigami for the 2μ Trs130 plasmid. This research was supported by grant GM-45444 from the National Institutes of Health (NIH) to N. Segev.

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Author notes

  1. Nadya Morozova and Yongheng Liang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biological Sciences, Laboratory for Molecular Biology, University of Illinois at Chicago, Molecular Biology Research Building, 900 South Ashland Avenue, Chicago, 60607, IL, USA

    Nadya Morozova, Yongheng Liang, Andrei A. Tokarev, Shu H. Chen, Randal Cox, Jelena Andrejic, Zhanna Lipatova & Nava Segev

  2. Department of Cellular and Molecular Medicine, University of California at San Diego, 9500 Gilman Drive, CMM-W318, La Jolla, 92093, CA, USA

    Vicki A. Sciorra & Scott D. Emr

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  1. Nadya Morozova
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Contributions

N.M., Y.L. and A.A.T. were responsible for the experimental work. S.H.C., J.A., Z.L., V.A.S. and S.D.E. contributed plasmids and strains. R.C. performed sequence analysis. N.S., with the help of the other authors, was involved in project planning, data analysis and writing.

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Correspondence to Nava Segev.

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The authors declare no competing financial interests.

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Supplementary Figures S1, S2, S3, S4, S5, S6 and Supplementary Table S1 (PDF 2769 kb)

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Morozova, N., Liang, Y., Tokarev, A. et al. TRAPPII subunits are required for the specificity switch of a Ypt–Rab GEF. Nat Cell Biol 8, 1263–1269 (2006). https://doi.org/10.1038/ncb1489

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