Journal Articles
The small GTP-binding protein rab6p is distributed from medial Golgi to the trans-Golgi network as determined by a confocal microscopic approach
C. Antony, C. Cibert, G. Geraud, A. Santa Maria, B. Maro, V. Mayau, B. Goud
Journal of Cell Science 1992 103: 785-796;

A key role in the regulation of membrane traffic is played by the rab proteins, members of a family of ras-related small GTP-binding proteins. This family comprises at least 25 identified members, the intracellular localization of only a few of which has been investigated. rab6p has been shown to be distributed along the exocytic pathway in association with the medial and trans regions of the Golgi apparatus. A confocal laser scanning microscopic (CLSM) approach coupled with image analysis was used to compare the localization of rab6p with selected reference Golgi markers by double immunofluorescence on culture cell lines. CLSM analysis shows that, under a set of well-defined conditions, one can investigate the possible colocalization of known markers of Golgi compartments and orientate a couple of labeled Golgi antigens with regard to the polarity of the Golgi apparatus. Thus, having validated the CLSM analysis, the localization of rab6p was studied and compared with some of these markers and the VSV-G protein in VSV (vesicular stomatitis virus)-infected cells blocked at 20 degrees C. rab6p is shown to be associated in all the cell lines used with the last cisternae of the Golgi apparatus and particularly with the trans-Golgi network (TGN), the site of protein sorting at the exit of the Golgi apparatus. These results were supported by an electron microscopic study using double-immunolabeled cryosections: rab6p was found in some flat cisternae of the Golgi stack and colocalized with the VSV-G protein in the TGN. Our results show that the small GTP-binding protein rab6p is distributed from medial Golgi to TGN along the exocytic pathway.

REFERENCES

    1. Bacon R. A.,
    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. Baker D.,
    2. Wuestehube L.,
    3. Schekman R.,
    4. Botstein D. and
    5. Segev N.
    (1990). GTP-binding Ypt1 protein and Ca2+ function independently in a cell-free protein transport reaction. Proc. Nat. Acad. Sci. USA 87, 355–359
    OpenUrlAbstract/FREE Full Text
    1. Baron M. D. and
    2. Garoff H.
    (1990). Mannosidase II and the 135K-DaGolgi-specific antigen recognized by monoclonal antibody 53FC3 are the same dimeric protein. J. Biol. Chem 265, 19928–19931
    OpenUrlAbstract/FREE Full Text
    1. Bourne H. R.
    (1988). Do GTPases direct membrane traffic in secretion?. Cell 53, 669–671
    OpenUrlCrossRefPubMedWeb of Science
    1. Burke B.,
    2. Griffiths G.,
    3. Reggio H.,
    4. Louvard D. and
    5. Warren G.
    (1982). A monoclonal antibody against a 135-k Golgi membrane protein. EMBO J 1, 1621–1628
    OpenUrlPubMedWeb of Science
    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. 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, 79–82
    OpenUrlCrossRefPubMed
    1. Gallwitz D.,
    2. Donath C. and
    3. Sander C.
    (1983). A yeast gene encoding a protein homologous to the human c-ras/bas proto-oncogene. Nature 306, 704–707
    OpenUrlCrossRefPubMed
    1. Gorvel J.-P.,
    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. and
    4. Novick P.
    (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
    OpenUrlCrossRefPubMedWeb of Science
    1. Griffiths G.,
    2. McDowall A.,
    3. Back R. and
    4. Dubochet J.
    (1984). On the preparation of cryosections for immunocytochemistry. J. Ultrastruct. Res 65, 78–89
    OpenUrl
    1. Griffiths G.,
    2. Pfeiffer S.,
    3. Simons K. and
    4. Matlin K.
    (1985). Exit of newly synthesized membrane proteins from the trans cisternae of the Golgi complex to the plasma membrane. J. Cell. Biol 101, 949–964
    OpenUrlAbstract/FREE Full Text
    1. Haubruck H.,
    2. Prange R.,
    3. Vorgias C. and
    4. Gallwitz D.
    (1989). The ras-related mouse ypt1 protein can functionally replace the YPT1 gene product in yeast. EMBO J 8, 1427–1432
    OpenUrlPubMedWeb of Science
    1. Jasmin B. J.,
    2. Goud B.,
    3. Camus G. and
    4. Cartaud J.
    (1992). The low molecular weight GTP-binding protein rab6p associates with distinct post-Golgi vesicles in Torpedo marmorata electrocytes. Neuroscience 4, 849–855
    OpenUrl
    1. Kreis T. E.
    (1986). Microinjected antibodies against the cytoplasmic domain of vesicular stomatitis virus glycoprotein block its transport to the cell surface. EMBO J 5, 931–941
    OpenUrlPubMedWeb of Science
    1. Lippincott-Schwarz J.,
    2. Yuan L.,
    3. Tipper C.,
    4. Amherdt M.,
    5. Orci L. and
    6. Klausner R. D.
    (1991). Brefeldin A's effects on endosomes, lysosomes and the TGN suggests a general mechanism for regulating organelle structure and membrane traffic. Cell 67, 601–616
    OpenUrlCrossRefPubMedWeb of Science
    1. Luzio J. P.,
    2. Brake B.,
    3. Banting G.,
    4. Howell K. E.,
    5. Braghetta P. and
    6. Stanley K. K.
    (1990). Identification, sequencing and expression of an integral membrane protein of the trans -Golgi network (TGN 38). Biochem. J 270, 97–102
    OpenUrlAbstract/FREE Full Text
    1. Matlin K. S. and
    2. Simons K.
    (1983). Reduced temperature prevents transfer of membrane glycoproteins to the cell surface but does not prevent terminal glycosylation. Cell 34, 233–243
    OpenUrlCrossRefPubMedWeb of Science
    1. Palade G.
    (1975). Intracellular aspects of the process of protein synthesis. Science 189, 347–358
    OpenUrlFREE Full Text
    1. Plutner H.,
    2. Cox A.,
    3. Pind S.,
    4. Khosravi-Far R.,
    5. Bourne J.,
    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. Reaves B. and
    2. Banting G.
    (1992). Perturbation of the Morphology of the trans -Golgi Network following Brefeldin A Treatment: Redistribution of a TGN-specific Integral Membrane Protein, TGN38. J. Cell Biol 116, 85–94
    OpenUrlAbstract/FREE Full Text
    1. Rothman J. E. and
    2. Orci L.
    (1992). Molecular dissection of the secretory pathway. Nature 355, 409–415
    OpenUrlCrossRefPubMedWeb of Science
    1. Salminen A. and
    2. Novick P.
    (1987). A ras-like protein is required for a post-Golgi event in yeast secretion. Cell 47, 527–538
    OpenUrl
    1. Segev N.
    (1991). Mediation of the attachment or fusion step in796vesicular 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. Touchot N.,
    2. Chardin P. and
    3. Tavitian A.
    (1987). Four additional members of the ras gene superfamily isolated by an oligonucleotide strategy: molecular cloning of YPT-related cDNAs from a rat brain library. Proc. Nat. Acad. Sci. USA 84, 8210–8214
    OpenUrlAbstract/FREE Full Text
    1. Walworth N. C.,
    2. Goud B.,
    3. Kabcenell A. and
    4. Novick P.
    (1989). Mutational analysis of SEC4 suggests a cyclical mechanism for the regulation of vesicular traffic. EMBO J 8, 1685–1693
    OpenUrlPubMedWeb of Science
    1. Yuan L.,
    2. Barriocanal J. G.,
    3. Bonifacino J. S. and
    4. Sandoval I. V.
    (1987). Two integral membrane proteins located in the cis-midlle and transport of the Golgi system acquire sialylated N-linked carbohydrates and display different turnovers and sensitivity to cAMP-dependent phosphorylation. J. Cell Biol 105, 215–227
    OpenUrlAbstract/FREE Full Text
    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
Back to top

 Download PDF

Email
The small GTP-binding protein rab6p is distributed from medial Golgi to the trans-Golgi network as determined by a confocal microscopic approach
C. Antony, C. Cibert, G. Geraud, A. Santa Maria, B. Maro, V. Mayau, B. Goud
Journal of Cell Science 1992 103: 785-796;
Permalink:
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Alerts

Article navigation

Other journals from The Company of Biologists

Cell scientist to watch – Kevin Corbett

Kevin Corbett

"[...]the most compelling science emerges when you put together a set of people with different expertise and outlooks on a problem.”

Kevin heads a research group at UC San Diego and the Ludwig Institute for Cancer Research, investigating meiosis and its links to carcinogenesis. He shares his thoughts on fostering successful collaborations and why momentum is key to getting a new lab up and running.

Cell science at a glance – Cellular functions of WASP family proteins at a glance

Regulation of F-actin generation at omegasome

In this poster and accompanying article, Olga Alekhina, Ezra Burstein and Daniel D. Billadeau provides insight into new functions of WASP family proteins from regulating the biogenesis of autophagosomes to recently identified roles in the nucleus.

Sticky wicket – Peerless II

Mole reflects on the criticisms of reviewers

Mole continues his philosophising on grant applications, this time delving into how to frame your project to capture the imaginations of the review panel.

Have you met our resident insectivore? Read more of his manic musings here.

Meeting report – Emerging concepts in cell organization

Collage of the 10th EMBO-Annaberg Workshop 2017

In January this year, the 10th Annaberg EMBO workshop was held in a medieval castle in Goldegg am See, Austria. David Teis and Wanda Kukulski report back on this lively exchange of the ideas and new concepts shaping our understanding of how cells are organized and how this organization impacts cellular functions.

Transfer to Biology Open

If your submission to Journal of Cell Science is unsuccessful, did you know you can transfer your paper and any reviews directly to our sister journal, Biology Open? The majority of papers transferred with reviews are accepted for publication. Find out how here.