Abstract
Rab GTPases control vesicle movement and tethering membrane events in membrane trafficking. We used the 38 human Rab GTPase activating proteins (GAPs) to identify which of the 60 Rabs encoded in the human genome function at the Golgi complex. Surprisingly, this screen identified only two GAPs, RN-tre and TBC1D20, disrupting both Golgi organization and protein transport. RN-tre is the GAP for Rab43, and controls retrograde transport into the Golgi from the endocytic pathway. TBC1D20 is the ER-localized GAP for Rab1, and is the only GAP blocking the delivery of secretory cargo from the ER to the cell surface. Strikingly, its expression causes the loss of the Golgi complex, highlighting the importance of Rab1 for Golgi biogenesis. These effects can be antagonized by reticulon, a binding partner for TBC1D20 in the ER. Together, these findings indicate that Rab1 and Rab43 are key Rabs required for the biogenesis and maintenance of a functional Golgi structure, and suggest that other Rabs acting at the Golgi complex are likely to be functionally redundant.
References
56
Referenced
176
-
Ahmadian, M. R., Stege, P., Scheffzek, K. and Wittinghofer, A. (1997). Confirmation of the arginine-finger hypothesis for the GAP-stimulated GTP-hydrolysis reaction of Ras. Nat. Struct. Biol.4, 686-689.
(
10.1038/nsb0997-686
) -
Albert, S. and Gallwitz, D. (1999). Two new members of a family of Ypt/Rab GTPase activating proteins. Promiscuity of substrate recognition. J. Biol. Chem.274, 33186-33189.
(
10.1074/jbc.274.47.33186
) -
Albert, S., Will, E. and Gallwitz, D. (1999). Identification of the catalytic domains and their functionally critical arginine residues of two yeast GTPase-activating proteins specific for Ypt/Rab transport GTPases. EMBO J.18, 5216-5225.
(
10.1093/emboj/18.19.5216
) -
Allan, B. B., Moyer, B. D. and Balch, W. E. (2000). Rab1 recruitment of p115 into a cis-SNARE complex: programming budding COPII vesicles for fusion. Science289, 444-448.
(
10.1126/science.289.5478.444
) -
Alvarez, C., Fujita, H., Hubbard, A. and Sztul, E. (1999). ER to Golgi transport: requirement for p115 at a pre-Golgi VTC stage. J. Cell Biol.147, 1205-1222.
(
10.1083/jcb.147.6.1205
) -
Bannykh, S. I., Rowe, T. and Balch, W. E. (1996). The organization of endoplasmic reticulum export complexes. J. Cell Biol.135, 19-35.
(
10.1083/jcb.135.1.19
) -
Barr, F. A., Preisinger, C., Kopajtich, R. and Korner, R. (2001). Golgi matrix proteins interact with p24 cargo receptors and aid their efficient retention in the Golgi apparatus. J. Cell Biol.155, 885-891.
(
10.1083/jcb.200108102
) -
Barrowman, J., Sacher, M. and Ferro-Novick, S. (2000). TRAPP stably associates with the Golgi and is required for vesicle docking. EMBO J.19, 862-869.
(
10.1093/emboj/19.5.862
) -
Bevis, B. J., Hammond, A. T., Reinke, C. A. and Glick, B. S. (2002). De novo formation of transitional ER sites and Golgi structures in Pichia pastoris. Nat. Cell Biol.4, 750-756.
(
10.1038/ncb852
) -
Bonfanti, L., Mironov, A. A., Jr, Martinez-Menarguez, J. A., Martella, O., Fusella, A., Baldassarre, M., Buccione, R., Geuze, H. J., Mironov, A. A. and Luini, A. (1998). Procollagen traverses the Golgi stack without leaving the lumen of cisternae: evidence for cisternal maturation. Cell95, 993-1003.
(
10.1016/S0092-8674(00)81723-7
) -
Bonifacino, J. S. and Glick, B. S. (2004). The mechanisms of vesicle budding and fusion. Cell116, 153-166.
(
10.1016/S0092-8674(03)01079-1
) -
Brennwald, P. and Novick, P. (1993). Interactions of three domains distinguishing the Ras-related GTP-binding proteins Ypt1 and Sec4. Nature362, 560-563.
(
10.1038/362560a0
) -
Cai, H., Yu, S., Menon, S., Cai, Y., Lazarova, D., Fu, C., Reinisch, K., Hay, J. C. and Ferro-Novick, S. (2007). TRAPPI tethers COPII vesicles by binding the coat subunit Sec23. Nature445, 941-944.
(
10.1038/nature05527
) -
Cao, X., Ballew, N. and Barlowe, C. (1998). Initial docking of ER-derived vesicles requires Uso1p and Ypt1p but is independent of SNARE proteins. EMBO J.17, 2156-2165.
(
10.1093/emboj/17.8.2156
) -
Connerly, P. L., Esaki, M., Montegna, E. A., Strongin, D. E., Levi, S., Soderholm, J. and Glick, B. S. (2005). Sec16 is a determinant of transitional ER organization. Curr. Biol.15, 1439-1447.
(
10.1016/j.cub.2005.06.065
) -
Cooper, A. A., Gitler, A. D., Cashikar, A., Haynes, C. M., Hill, K. J., Bhullar, B., Liu, K., Xu, K., Strathearn, K. E., Liu, F. et al. (2006). Alpha-synuclein blocks ER-Golgi traffic and Rab1 rescues neuron loss in Parkinson's models. Science313, 324-328.
(
10.1126/science.1129462
) -
De Antoni, A., Schmitzova, J., Trepte, H. H., Gallwitz, D. and Albert, S. (2002). Significance of GTP hydrolysis in Ypt1p-regulated endoplasmic reticulum to Golgi transport revealed by the analysis of two novel Ypt1-GAPs. J. Biol. Chem.277, 41023-41031.
(
10.1074/jbc.M205783200
) -
De Craene, J. O., Coleman, J., Estrada de Martin, P., Pypaert, M., Anderson, S., Yates, J. R., 3rd, Ferro-Novick, S. and Novick, P. (2006). Rtn1p is involved in structuring the cortical endoplasmic reticulum. Mol. Biol. Cell17, 3009-3020.
(
10.1091/mbc.e06-01-0080
) -
Fuchs, E., Haas, A. K., Spooner, R. A., Yoshimura, S., Lord, J. M. and Barr, F. A. (2007). Specific Rab GTPase-activating proteins define the Shiga toxin and epidermal growth factor uptake pathways. J. Cell Biol.177, 1133-1143.
(
10.1083/jcb.200612068
) -
Glick, B. S. (2002). Can the Golgi form de novo? Nat. Rev. Mol. Cell Biol.3, 615-619.
(
10.1038/nrm877
) -
Guo, Y. and Linstedt, A. D. (2006). COPII-Golgi protein interactions regulate COPII coat assembly and Golgi size. J. Cell Biol.174, 53-63.
(
10.1083/jcb.200604058
) -
Haas, A. K., Fuchs, E., Kopajtich, R. and Barr, F. A. (2005). A GTPase-activating protein controls Rab5 function in endocytic trafficking. Nat. Cell Biol.7, 887-893.
(
10.1038/ncb1290
) -
Jena, B. P., Brennwald, P., Garrett, M. D., Novick, P. and Jamieson, J. D. (1992). Distinct and specific GAP activities in rat pancreas act on the yeast GTP-binding proteins Ypt1 and Sec4. FEBS Lett.309, 5-9.
(
10.1016/0014-5793(92)80727-X
) -
Jones, S., Newman, C., Liu, F. and Segev, N. (2000). The TRAPP complex is a nucleotide exchanger for Ypt1 and Ypt31/32. Mol. Biol. Cell11, 4403-4411.
(
10.1091/mbc.11.12.4403
) -
Losev, E., Reinke, C. A., Jellen, J., Strongin, D. E., Bevis, B. J. and Glick, B. S. (2006). Golgi maturation visualized in living yeast. Nature441, 1002-1006.
(
10.1038/nature04717
) -
Matsuura-Tokita, K., Takeuchi, M., Ichihara, A., Mikuriya, K. and Nakano, A. (2006). Live imaging of yeast Golgi cisternal maturation. Nature441, 1007-1010.
(
10.1038/nature04737
) -
Mironov, A. A., Weidman, P. and Luini, A. (1997). Variations on the intracellular transport theme: maturing cisternae and trafficking tubules. J. Cell Biol.138, 481-484.
(
10.1083/jcb.138.3.481
) -
Mironov, A. A., Mironov, A. A., Jr, Beznoussenko, G. V., Trucco, A., Lupetti, P., Smith, J. D., Geerts, W. J., Koster, A. J., Burger, K. N., Martone, M. E. et al. (2003). ER-to-Golgi carriers arise through direct en bloc protrusion and multistage maturation of specialized ER exit domains. Dev. Cell5, 583-594.
(
10.1016/S1534-5807(03)00294-6
) -
Morsomme, P. and Riezman, H. (2002). The Rab GTPase Ypt1p and tethering factors couple protein sorting at the ER to vesicle targeting to the Golgi apparatus. Dev. Cell2, 307-317.
(
10.1016/S1534-5807(02)00133-8
) -
Munro, S. (2002). Organelle identity and the targeting of peripheral membrane proteins. Curr. Opin. Cell Biol.14, 506-514.
(
10.1016/S0955-0674(02)00350-2
) -
Pan, X., Eathiraj, S., Munson, M. and Lambright, D. G. (2006). TBC-domain GAPs for Rab GTPases accelerate GTP hydrolysis by a dual-finger mechanism. Nature442, 303-306.
(
10.1038/nature04847
) -
Pelletier, L., Stern, C. A., Pypaert, M., Sheff, D., Ngo, H. M., Roper, N., He, C. Y., Hu, K., Toomre, D., Coppens, I. et al. (2002). Golgi biogenesis in Toxoplasma gondii. Nature418, 548-552.
(
10.1038/nature00946
) -
Peter, F., Nuoffer, C., Pind, S. N. and Balch, W. E. (1994). Guanine nucleotide dissociation inhibitor is essential for Rab1 function in budding from the endoplasmic reticulum and transport through the Golgi stack. J. Cell Biol.126, 1393-1406.
(
10.1083/jcb.126.6.1393
) -
Pfeffer, S. R. (1999). Transport-vesicle targeting: tethers before SNAREs. Nat. Cell Biol.1, E17-E22.
(
10.1038/8967
) -
Pfeffer, S. and Aivazian, D. (2004). Targeting Rab GTPases to distinct membrane compartments. Nat. Rev. Mol. Cell Biol.5, 886-896.
(
10.1038/nrm1500
) -
Puthenveedu, M. A., Bachert, C., Puri, S., Lanni, F. and Linstedt, A. D. (2006). GM130 and GRASP65-dependent lateral cisternal fusion allows uniform Golgi-enzyme distribution. Nat. Cell Biol.8, 238-248.
(
10.1038/ncb1366
) -
Rak, A., Fedorov, R., Alexandrov, K., Albert, S., Goody, R. S., Gallwitz, D. and Scheidig, A. J. (2000). Crystal structure of the GAP domain of Gyp1p: first insights into interaction with Ypt/Rab proteins. EMBO J.19, 5105-5113.
(
10.1093/emboj/19.19.5105
) -
Richardson, C. J., Jones, S., Litt, R. J. and Segev, N. (1998). GTP hydrolysis is not important for Ypt1 GTPase function in vesicular transport. Mol. Cell. Biol.18, 827-838.
(
10.1128/MCB.18.2.827
) -
Sacher, M., Barrowman, J., Wang, W., Horecka, J., Zhang, Y., Pypaert, M. and Ferro-Novick, S. (2001). TRAPP I implicated in the specificity of tethering in ER-to-Golgi transport. Mol. Cell7, 433-442.
(
10.1016/S1097-2765(01)00190-3
) -
Segev, N. (1991). Mediation of the attachment or fusion step in vesicular transport by the GTP-binding Ypt1 protein. Science252, 1553-1556.
(
10.1126/science.1904626
) -
Shima, D. T., Haldar, K., Pepperkok, R., Watson, R. and Warren, G. (1997). Partitioning of the Golgi apparatus during mitosis in living HeLa cells. J. Cell Biol.137, 1211-1228.
(
10.1083/jcb.137.6.1211
) -
Short, B., Preisinger, C., Korner, R., Kopajtich, R., Byron, O. and Barr, F. A. (2001). A GRASP55-rab2 effector complex linking Golgi structure to membrane traffic. J. Cell Biol.155, 877-883.
(
10.1083/jcb.200108079
) -
Short, B., Haas, A. and Barr, F. A. (2005). Golgins and GTPases, giving identity and structure to the Golgi apparatus. Biochim. Biophys. Acta1744, 383-395.
(
10.1016/j.bbamcr.2005.02.001
) -
Shorter, J., Watson, R., Giannakou, M. E., Clarke, M., Warren, G. and Barr, F. A. (1999). GRASP55, a second mammalian GRASP protein involved in the stacking of Golgi cisternae in a cell-free system. EMBO J.18, 4949-4960.
(
10.1093/emboj/18.18.4949
) -
Strom, M., Vollmer, P., Tan, T. J. and Gallwitz, D. (1993). A yeast GTPase-activating protein that interacts specifically with a member of the Ypt/Rab family. Nature361, 736-739.
(
10.1038/361736a0
) -
Tisdale, E. J. and Balch, W. E. (1996). Rab2 is essential for the maturation of pre-Golgi intermediates. J. Biol. Chem.271, 29372-29379.
(
10.1074/jbc.271.46.29372
) -
Tisdale, E. J., Bourne, J. R., Khosravi-Far, R., Der, C. J. and Balch, W. E. (1992). GTP-binding mutants of rab1 and rab2 are potent inhibitors of vesicular transport from the endoplasmic reticulum to the Golgi complex. J. Cell Biol.119, 749-761.
(
10.1083/jcb.119.4.749
) -
Vetter, I. R. and Wittinghofer, A. (2001). The guanine nucleotide-binding switch in three dimensions. Science294, 1299-1304.
(
10.1126/science.1062023
) -
Voeltz, G. K., Prinz, W. A., Shibata, Y., Rist, J. M. and Rapoport, T. A. (2006). A class of membrane proteins shaping the tubular endoplasmic reticulum. Cell124, 573-586.
(
10.1016/j.cell.2005.11.047
) -
Wakana, Y., Koyama, S., Nakajima, K., Hatsuzawa, K., Nagahama, M., Tani, K., Hauri, H. P., Melancon, P. and Tagaya, M. (2005). Reticulon 3 is involved in membrane trafficking between the endoplasmic reticulum and Golgi. Biochem. Biophys. Res. Commun.334, 1198-1205.
(
10.1016/j.bbrc.2005.07.012
) -
Wang, W., Sacher, M. and Ferro-Novick, S. (2000). TRAPP stimulates guanine nucleotide exchange on Ypt1p. J. Cell Biol.151, 289-296.
(
10.1083/jcb.151.2.289
) -
Waters, M. G. and Hughson, F. M. (2000). Membrane tethering and fusion in the secretory and endocytic pathways. Traffic1, 588-597.
(
10.1034/j.1600-0854.2000.010802.x
) -
Watson, P., Townley, A. K., Koka, P., Palmer, K. J. and Stephens, D. J. (2006). Sec16 defines endoplasmic reticulum exit sites and is required for secretory cargo export in mammalian cells. Traffic7, 1678-1687.
(
10.1111/j.1600-0854.2006.00493.x
) -
Weide, T., Teuber, J., Bayer, M. and Barnekow, A. (2003). MICAL-1 isoforms, novel rab1 interacting proteins. Biochem. Biophys. Res. Commun.306, 79-86.
(
10.1016/S0006-291X(03)00918-5
) -
Yu, S., Satoh, A., Pypaert, M., Mullen, K., Hay, J. C. and Ferro-Novick, S. (2006). mBet3p is required for homotypic COPII vesicle tethering in mammalian cells. J. Cell Biol.174, 359-368.
(
10.1083/jcb.200603044
) -
Zerial, M. and McBride, H. (2001). Rab proteins as membrane organizers. Nat. Rev. Mol. Cell Biol.2, 107-117.
(
10.1038/35052055
)
Dates
Type | When |
---|---|
Created | 18 years ago (Aug. 7, 2007, 9:54 p.m.) |
Deposited | 4 years, 4 months ago (April 25, 2021, 4:53 p.m.) |
Indexed | 3 days, 21 hours ago (Aug. 23, 2025, 9:55 p.m.) |
Issued | 17 years, 11 months ago (Sept. 1, 2007) |
Published | 17 years, 11 months ago (Sept. 1, 2007) |
Published Print | 17 years, 11 months ago (Sept. 1, 2007) |
@article{Haas_2007, title={Analysis of GTPase-activating proteins: Rab1 and Rab43 are key Rabs required to maintain a functional Golgi complex in human cells}, volume={120}, ISSN={0021-9533}, url={http://dx.doi.org/10.1242/jcs.014225}, DOI={10.1242/jcs.014225}, number={17}, journal={Journal of Cell Science}, publisher={The Company of Biologists}, author={Haas, Alexander K. and Yoshimura, Shin-ichiro and Stephens, David J. and Preisinger, Christian and Fuchs, Evelyn and Barr, Francis A.}, year={2007}, month=sep, pages={2997–3010} }