Journal Articles
Rab6 is associated with a compartment that transports rhodopsin from the trans-Golgi to the site of rod outer segment disk formation in frog retinal photoreceptors
D. Deretic, D.S. Papermaster
Journal of Cell Science 1993 106: 803-813;

Summary

The biogenesis of light sensitive membranes in retinal rod photoreceptors involves polarized sorting and targeting of newly synthesized rhodopsin to a specialized domain, the rod outer segment (ROS). We have isolated and characterized the population of post-Golgi membranes that mediate intracellular transport of rhodopsin. In the present study we have examined the association of small (20-25 kDa) GTP-binding (G) proteins with these membranes. We found that one of the small G proteins, rab6, behaves like an integral membrane protein of the post-Golgi vesicles, although approximately 30% of rab6 is soluble. The distribution of the membrane-associated and the soluble forms is highly polarized. By confocal and EM immunocytochemistry it can be seen that most of rab6 is associated with the photoreceptor trans-Golgi cisternae, trans-Golgi network (TGN) and post-Golgi vesicles. The photoreceptor axon and synaptic terminal are unlabeled, but dendrites of deeper retinal layers are labeled. The distribution of rab6 across sucrose density gradient fractions parallels the distribution of sialyltransferase (a TGN marker) activity. About 9% of membrane-bound rab6 is associated, however, with the rhodopsin-bearing sialyltransferase-free post-Golgi vesicles, which represent a very small fraction (< 1%) of the total retinal membranes. Rab6 is absent from the mature ROS disk membranes but it is present at the sites of new ROS disk formation and in the ROS cytoplasm. This suggests that rab6 becomes soluble upon disk membrane formation. Therefore, rab6 may function not only as a component of the sorting machinery of photoreceptors that delivers rhodopsin to its appropriate subcellular domain but may also participate in some aspects of ROS disk morphogenesis.

REFERENCES

    1. Antony C.,
    2. Cibert C.,
    3. Geraud G.,
    4. Santa Maria A.,
    5. Maro B.,
    6. Mayau V. and
    7. Goud B.
    (1992). The small GTP-binding protein rab6p is distributed from medial Golgi to the trans -Golgi network as determined by a confocal microscopic approach. J. Cell Sci 103, 785–796
    OpenUrlAbstract/FREE Full Text
    1. Araki S.,
    2. Kikuchi A.,
    3. Hata Y.,
    4. Isomura M. and
    5. Takai Y.
    (1990). Regulation of reversible bind of smg p25A, a ras p21-like GTP-binding protein, to synaptic plasma membranes and vesicles by its specific regulatory protein, GDP dissociation inhibitor. J. Biol. Chem 265, 13007–13015
    OpenUrlAbstract/FREE Full Text
    1. Bacon R.,
    2. Salminen A.,
    3. Ruohola H.,
    4. Novick P. and
    5. Ferro-Novick S.
    (1989). The GTP-binding protein Ypt1 is required for transport in vitro: the Golgi apparatus is defective in Ypt1 mutants. J. Cell Biol 109, 1015–1022
    OpenUrlAbstract/FREE Full Text
    1. Balch W. E.
    (1990). Small GTP-binding proteins in vesicular transport. Trends Biochem. Sci 15, 473–477
    OpenUrlCrossRefPubMedWeb of Science
    1. Berryman M. A. and
    2. Rodewald R. D.
    (1990). An enhanced method for post-embedding immunocytochemical staining which preserves cell membranes. J. Histochem. Cytochem 38, 159–170
    OpenUrlAbstract/FREE Full Text
    1. Brandli A.,
    2. Hansson G.,
    3. Rodriguez-Boulan E. and
    4. Simons K.
    (1988). A polarized epithelial cell mutant deficient in translocation of UDP-galactose into the Golgi complex. J. Biol. Chem 263, 16283–16290
    OpenUrlAbstract/FREE Full Text
    1. Bucci C.,
    2. Parton R.,
    3. Mather I.,
    4. Stunnenberg H.,
    5. Simons K.,
    6. Hoflack B. and
    7. Zerial M.
    (1992). The small GTPase rab5 functions as a regulatory factor in the early endocytic pathway. Cell 70, 715–728
    OpenUrlCrossRefPubMedWeb of Science
    1. Burger P. M.,
    2. Mehl E.,
    3. Cameron P. L.,
    4. Maycox P. R.,
    5. Baumert M.,
    6. Lottspeich F.,
    7. DeCamilli P. and
    8. Jahn R.
    (1989). Synaptic vesicles immunoisolated from rat cerebral cortex contain high levels of glutamate. Neuron 3, 715–720
    OpenUrlCrossRefPubMedWeb of Science
    1. Chaitin M. H. and
    2. Burnside B.
    (1989). Actin filament polarity at the site of rod outer segment disk morphogenesis. Invest. Ophthalmol. Vis. Sci 30, 2461–2469
    OpenUrlAbstract/FREE Full Text
    1. Chaitin M.,
    2. Schneider B.,
    3. Hall M. and
    4. Papermaster D.
    (1984). Actin in the photoreceptor connecting cilium: immunocytochemical localization to the site of outer segment disk formation. J. Cell Biol 99, 239–247
    OpenUrlAbstract/FREE Full Text
    1. Chavrier P.,
    2. Parton R. G.,
    3. Hauri H.-P.,
    4. Simons K. and
    5. Zerial M.
    (1990). Localization of low molecular weight GTP-binding proteins to exocytic and endocytic compartments. Cell 62, 317–329
    OpenUrlCrossRefPubMedWeb of Science
    1. Chavrier P.,
    2. Vingron M.,
    3. Sander C.,
    4. Simons K. and
    5. Zerial M.
    (1990). Molecular cloning of YPT1/SEC4 -related cDNAs from an epithelial cell line. Mol. Cell. Biol 10, 6578–6585
    OpenUrlAbstract/FREE Full Text
    1. Chavrier P.,
    2. Gorvel J. P.,
    3. Stelzer E.,
    4. Simons K.,
    5. Gruenberg J. and
    6. Zerial M.
    (1991). Hypervariable C-terminal domain of rab proteins acts as a targeting signal. Nature 353, 769–772
    OpenUrlCrossRefPubMedWeb of Science
    1. Darchen F.,
    2. Zahraoui A.,
    3. Hammel F.,
    4. Monteils M. P.,
    5. Tavitian A. and
    6. Scherman D.
    (1990). Association of the GTP-binding protein Rab3A with bovine adrenal chromaffin granules. Proc. Nat. Acad. Sci. USA 87, 5692–5696
    OpenUrlAbstract/FREE Full Text
    1. Deretic D. and
    2. Papermaster D. S.
    (1991). Polarized sorting of rhodopsin on post-Golgi membranes in frog retinal photoreceptor cells. J. Cell Biol 113, 1281–1293
    OpenUrlAbstract/FREE Full Text
    1. Dunn W. A. and
    2. Hubbard A. L.
    (1984). Receptor-mediated endocytosis of epidermal growth factor by hepatocytes in the perfused rat liver: ligand and receptor dynamics. J. Cell Biol 98, 2148–2159
    OpenUrlAbstract/FREE Full Text
    1. Fischer von Mollard G.,
    2. Mignery G. A.,
    3. Baumert M.,
    4. Perin M. S.,
    5. Hanson T. J.,
    6. Burger P. M.,
    7. Jahn R. and
    8. Sudhof T. C.
    (1990). Rab3 is a small GTP-binding protein exclusively localized to synaptic vesicles. Proc. Nat. Acad. Sci. USA 87, 1988–1992
    OpenUrlAbstract/FREE Full Text
    1. Fischer von Mollard G.,
    2. Sudhof T. C. and
    3. Jahn R.
    (1991). A small GTP-binding protein dissociates from synaptic vesicles during exocytosis. Nature 349, 79–81
    OpenUrlCrossRefPubMed
    1. Gorvel J.,
    2. Chavrier P.,
    3. Zerial M. and
    4. Gruenberg J.
    (1991). Rab5 controls early endosome fusion in vitro. Cell 64, 915–925
    OpenUrlCrossRefPubMedWeb of Science
    1. Goud B.,
    2. Salminen A.,
    3. Walworth N. C. and
    4. Novick P. J.
    (1988). A GTP-binding protein required for secretion rapidly associates with secretory vesicles and the plasma membrane in yeast. Cell 53, 753–768
    OpenUrlCrossRefPubMedWeb of Science
    1. Goud B.,
    2. Zahraoui A.,
    3. Tavitian A. and
    4. Saraste J.
    (1990). Small GTP-binding protein associated with Golgi cisternae. Nature 345, 553–556
    OpenUrlCrossRefPubMed
    1. Goud B. and
    2. McCaffrey M.
    (1991). Small GTP-binding proteins and their role in transport. Curr. Opin. Cell Biol 3, 626–633
    OpenUrlCrossRefPubMed
    1. Goud B.
    (1992). Small GTP-binding proteins as compartmental markers. Semin. Cell Biol 3, 301–307
    OpenUrlCrossRefPubMed
    1. Griffiths G. and
    2. Simons K.
    (1986). The trans -Golgi network: sorting at the exit site of the Golgi complex. Science 234, 438–443
    OpenUrlAbstract/FREE Full Text
    1. Hall A.
    (1990). The cellular functions of small GTP-binding proteins. Science 249, 635–640
    OpenUrlAbstract/FREE Full Text
    1. Jasmin B.,
    2. Goud B.,
    3. Camus G. and
    4. Cartaud J.
    (1992). The low molecular weight guanosine triphosphate-binding protein rab6p associates with distinct post-Golgi vesicles in Torpedo marmorata electrocytes. Neuroscience 49, 849–855
    OpenUrlCrossRefPubMedWeb of Science
    1. Khosravi-Far R.,
    2. Lutz R. J.,
    3. Cox A. D.,
    4. Conroy L.,
    5. Bourne J. R.,
    6. Sinensky M.,
    7. Balch W. E.,
    8. Buss J. E. and
    9. Der C. J.
    (1991). Isoprenoid modification of rab proteins terminating in CC or CXC motifs. Proc. Nat. Acad. Sci. USA 88, 6264–6268
    OpenUrlAbstract/FREE Full Text
    1. Kinsella B. T. and
    2. Maltese W. A.
    (1992). Rab GTP-binding proteins with three different carboxyl-terminal cysteine motifs are modified in vivo by 20-carbon isoprenoids. J. Biol. Chem 267, 3940–3945
    OpenUrlAbstract/FREE Full Text
    1. Laemmli U. K.
    (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685
    OpenUrlCrossRefPubMedWeb of Science
    1. Lapetina E. G. and
    2. Reep B. R.
    (1987). Specific binding of [- 32P]GTP to cytosolic and membrane-bound proteins of human platelets correlates with the activation of phospholipase. C. Proc. Nat. Acad. Sci. USA 84, 2261–2265
    OpenUrlAbstract/FREE Full Text
    1. Lombardi D.,
    2. Soldati T.,
    3. Riederer M. A.,
    4. Goda Y.,
    5. Zerial M. and
    6. Pfeffer S. R.
    (1993). Rab9 functions in transport between late endosomes and the trans- Golgi network. EMBO J 12, 677–682
    OpenUrlPubMedWeb of Science
    1. Matsudaira P.
    (1987). Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J. Biol. Chem 262, 10035–10038
    OpenUrlAbstract/FREE Full Text
    1. Matteoli M.,
    2. Takei K.,
    3. Cameron R.,
    4. Hurlbut P.,
    5. Johnson P. A.,
    6. Sudhof T. C.,
    7. Jahn R. and
    8. De Camilli P.
    (1991). Association of rab 3a with synaptic vesicles at late stages of the secretory pathway. J. Cell Biol 115, 625–633
    OpenUrlAbstract/FREE Full Text
    1. Nakano A. and
    2. Muramatsu M.
    (1989). A novel GTP-binding protein, Sar 1p, is involved in transport from the endoplasmic reticulum to the Golgi apparatus. J. Cell Biol 109, 2677–2691
    OpenUrlAbstract/FREE Full Text
    1. Nilsson T.,
    2. Pypaert M.,
    3. Hoe M.,
    4. Slusarewicz P.,
    5. Berger E. and
    6. Warren G.
    (1993). Overlapping distribution of two glycosyltransferases in the Golgi apparatus of HeLa cells. J. Cell Biol 120, 5–13
    OpenUrlAbstract/FREE Full Text
    1. Papermaster D. S. and
    2. Dreyer W. J.
    (1974). Rhodopsin content in the outer segment membranes of bovine and frog retinal rods. Biochemistry 13, 2438–2444
    OpenUrlCrossRefPubMed
    1. Papermaster D. S.,
    2. Schneider B. G.,
    3. Defoe D. and
    4. Besharse J. C.
    (1986). Biosynthesis and vectorial transport of opsin on vesicles in retinal rod photoreceptors. J. Histochem. Cytochem 34, 5–16
    OpenUrlAbstract/FREE Full Text
    1. Peters K.-R.,
    2. Palade G. E.,
    3. Schneider B. G. and
    4. Papermaster D. S.
    (1983). Fine structure of a periciliary ridge complex of frog retinal rod cells revealed by ultrahigh resolution scanning electron microscopy. J. Cell Biol 96, 265–276
    OpenUrlAbstract/FREE Full Text
    1. Plutner H.,
    2. Cox A.,
    3. Pind S.,
    4. Khosravi-Far R.,
    5. Bourne J. R.,
    6. Schwaninger R.,
    7. Der C. and
    8. Balch W.
    (1991). Rab1b regulates vesicular transport between the endoplasmic reticulum and successive Golgi compartments. J. Cell Biol 115, 31–43
    OpenUrlAbstract/FREE Full Text
    1. Regazzi R.,
    2. Kikuchi A.,
    3. Takai Y. and
    4. Wollheim C. B.
    (1992). The small GTP-binding proteins in the cytosol of insulin-secreting cells are complexed to GDP dissociation inhibitor proteins. J. Biol. Chem 267, 17512–17519
    OpenUrlAbstract/FREE Full Text
    1. Regnier-Vigouroux A.,
    2. Tooze S. and
    3. Huttner W.
    (1991). Newly synthesized synaptophysin is transported to synaptic-like microvesicles via constitutive secretory vesicles and the plasma membrane. EMBO J 10, 3589–3601
    OpenUrlPubMedWeb of Science
    1. Rexach M. F. and
    2. Schekman R. W.
    (1991). Distinct biochemical requirements for the budding, targeting and fusion of ER-derived transport vesicles. J. Cell Biol 114, 219–229
    OpenUrlAbstract/FREE Full Text
    1. Roth J.,
    2. Taatjes D.,
    3. Lucocq J.,
    4. Weinstein J. and
    5. Paulson J.
    (1985). Demonstration of an extensive trans -tubular network continuous with the Golgi apparatus stack that may function in glycosylation. Cell 43, 287–295
    OpenUrlCrossRefPubMedWeb of Science
    1. Salminen A. and
    2. Novick P. J.
    (1987). A ras -like protein is required for a post-Golgi event in yeast secretion. Cell 49, 527–538
    OpenUrlCrossRefPubMedWeb of Science
    1. Schmied R. and
    2. Holtzman E.
    (1989). Involvement of the Golgi apparatus in sorting of materials to opposite ends of frog rod retinal photoreceptors. J. Neurobiol 20, 115–138
    OpenUrlCrossRefPubMed
    1. Seabra M.,
    2. Goldstein J.,
    3. Sudhof T. and
    4. Brown M.
    (1992). Rab geranylgeranyl transferase- a multisubunit enzyme that prenylates GTP-binding proteins terminating in Cys-X-Cys or Cys-Cys. J. Biol. Chem 267, 14497–14503
    OpenUrlAbstract/FREE Full Text
    1. Seabra M.,
    2. Brown M.,
    3. Slaughter C.,
    4. Sudhof T. and
    5. Goldstein J.
    (1992). Purification of component A of Rab geranylgeranyl transferase: Possible identity with the choroideremia gene product. Cell 70, 1049–1057
    OpenUrlCrossRefPubMedWeb of Science
    1. Seabra M.,
    2. Brown M. and
    3. Goldstein J.
    (1993). Retinal degeneration in choroideremia: deficiency of Rab geranylgeranyl transferase. Science 259, 377–381
    OpenUrlAbstract/FREE Full Text
    1. Segev N.
    (1991). Mediation of the attachment or fusion step in vesicular transport by the GTP-binding Ypt1 protein. Science 252, 1553–1556
    OpenUrlAbstract/FREE Full Text
    1. Segev N.,
    2. Mulholland J. and
    3. Botstein D.
    (1988). The yeast GTP-binding YPT1 protein and a mammalian counterpart are associated with the secretion machinery. Cell 52, 915–924
    OpenUrlCrossRefPubMedWeb of Science
    1. Shirataki H.,
    2. Kaibuchi K.,
    3. Hiroyoshi M.,
    4. Isomura M.,
    5. Araki S.,
    6. Sasaki T. and
    7. Takai Y.
    (1991). Inhibition of the action of the stimulatory GDP/GTP exchange protein for smg p21 by the geranylgeranylated synthetic peptides designed from its C-terminal region. J. Biol. Chem 266, 20672–20677
    OpenUrlAbstract/FREE Full Text
    1. Steinberg R. H.,
    2. Fisher S. K. and
    3. Anderson D. H.
    (1980). Disc morphogenesis in vertebrate photoreceptors. J. Comp. Neurol 190, 501–518
    OpenUrlCrossRefPubMedWeb of Science
    1. Takai Y.,
    2. Kaibuchi K.,
    3. Kikuchi A. and
    4. Kawata M.
    (1992). Small GTP-binding proteins. Int. Rev. Cytol 133, 187–230
    OpenUrlCrossRefPubMedWeb of Science
    1. Valencia A.,
    2. Chardin P.,
    3. Wittinghofer A. and
    4. Sander C.
    (1991). The ras protein family: evolutionary tree and role of conserved amino acids. Biochemistry 30, 4637–4848
    OpenUrlCrossRefPubMed
    1. van der Sluijs P.,
    2. Hull M.,
    3. Webster P.,
    4. Male P.,
    5. Goud B. and
    6. Mellman I.
    (1992). The small GTP-binding protein rab4 controls an early sorting event on the endocytic pathway. Cell 70, 792–740
    OpenUrl
    1. Walworth N. C.,
    2. Goud B.,
    3. Kabcenell A. K. and
    4. Novick P. J.
    (1989). Mutational analysis of SEC4 suggests a cyclical mechanism for the regulation of vesicular traffic. EMBO J 8, 1685–1693
    OpenUrlPubMedWeb of Science
    1. Wandinger-Ness A.,
    2. Bennet M. K.,
    3. Antony C. and
    4. Simons K.
    (1990). Distinct transport vesicles mediate delivery of plasma membrane proteins to the apical and basolateral domains of MDCK cells. J. Cell Biol 111, 987–1000
    OpenUrlAbstract/FREE Full Text
    1. Yang C.,
    2. Mayau V.,
    3. Godeau F. and
    4. Goud B.
    (1992). Characterization of the unprocessed and processed forms of rab6 expressed in baculovirus/insect cell systems. Biochem. Biophys. Res. Commun 182, 1499–1505
    OpenUrlCrossRefPubMed
    1. Zahraoui A.,
    2. Touchot N.,
    3. Chardin P. and
    4. Tavitian A.
    (1989). The human rab genes encode a family of GTP-binding proteins related to yeast YPT1 and SEC4 products involved in secretion. J. Biol. Chem 264, 12394–12401
    OpenUrlAbstract/FREE Full Text
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Rab6 is associated with a compartment that transports rhodopsin from the trans-Golgi to the site of rod outer segment disk formation in frog retinal photoreceptors
D. Deretic, D.S. Papermaster
Journal of Cell Science 1993 106: 803-813;
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