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Science 11 October 1996:
Vol. 274. no. 5285, pp. 239 - 242
DOI: 10.1126/science.274.5285.239
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Reports

Role of GTP Hydrolysis in Fission of Caveolae Directly from Plasma Membranes

Jan E. Schnitzer, * Phil Oh, Deirdre P. McIntosh

Caveolae are specialized invaginated cell surface microdomains of undefined function. A cell-free system that reconstituted fission of caveolae from lung endothelial plasma membranes was developed. Addition of cytosol and the hydrolysis of guanosine triphosphate (GTP) induced caveolar fission. The budded caveolae were isolated as vesicles rich in caveolin and the sialoglycolipid GM1 but not glycosyl-phosphatidylinositol (GPI)-anchored proteins. These vesicles contained the molecular machinery for endocytosis and transcytosis. In permeabilized endothelial cells, GTP stimulated, whereas GTPgamma S prevented, caveolar budding and endocytosis of the cholera toxin B chain to endosomes. Thus, caveolae may bud to form discrete carrier vesicles that participate in membrane trafficking.

Department of Pathology, Harvard Medical School, Beth Israel Hospital, Research North, 99 Brookline Avenue, Boston, MA 02215, USA.
*   To whom correspondence should be addressed. E-mail: jschnitz{at}bih.harvard.edu

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Direct demonstration of the endocytic function of caveolae by a cell-free assay.
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M. G. Waugh, D. Lawson, S. K. Tan, and J. J. Hsuan (1998)
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Ganglioside Structure Dictates Signal Transduction by Cholera Toxin and Association with Caveolae-like Membrane Domains in Polarized Epithelia.
A. A. Wolf, M. G. Jobling, S. Wimer-Mackin, M. Ferguson-Maltzman, J. L. Madara, R. K. Holmes, and W. I. Lencer (1998)
J. Cell Biol. 141, 917-927
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Dynamin-mediated Internalization of Caveolae.
J. R. Henley, E. W.A. Krueger, B. J. Oswald, and M. A. McNiven (1998)
J. Cell Biol. 141, 85-99
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Dynamin at the Neck of Caveolae Mediates Their Budding to Form Transport Vesicles by GTP-driven Fission from the Plasma Membrane of Endothelium.
P. Oh, D. P. McIntosh, and J. E. Schnitzer (1998)
J. Cell Biol. 141, 101-114
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Functional association between Arf and RalA in active phospholipase D complex.
J.-Q. Luo, X. Liu, P. Frankel, T. Rotunda, M. Ramos, J. Flom, H. Jiang, L. A. Feig, A. J. Morris, R. A. Kahn, et al. (1998)
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Basolateral Distribution of Caveolin-1 in the Kidney: Absence from H+-ATPase-coated Endocytic Vesicles in Intercalated Cells.
S. Breton, M. P. Lisanti, R. Tyszkowski, M. McLaughlin, and D. Brown (1998)
J. Histochem. Cytochem. 46, 205-214
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Endocytic internalization in yeast and animal cells: similar and different.
M. Geli and H Riezman (1998)
J. Cell Sci. 111, 1031-1037
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Expression of caveolin-1 and polarized formation of invaginated caveolae in Caco-2 and MDCK II cells.
U Vogel, K Sandvig, and B van Deurs (1998)
J. Cell Sci. 111, 825-832
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Gp60 Activation Mediates Albumin Transcytosis in Endothelial Cells by Tyrosine Kinase-dependent Pathway.
C. Tiruppathi, W. Song, M. Bergenfeldt, P. Sass, and A. B. Malik (1997)
J. Biol. Chem. 272, 25968-25975
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Insulin-stimulated Tyrosine Phosphorylation of Caveolin Is Specific for the Differentiated Adipocyte Phenotype in 3T3-L1 Cells.
C. C. Mastick and A. R. Saltiel (1997)
J. Biol. Chem. 272, 20706-20714
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The First 35 Amino Acids and Fatty Acylation Sites Determine the Molecular Targeting of Endothelial Nitric Oxide Synthase into the Golgi Region of Cells: A Green Fluorescent Protein Study.
J. Liu, T. E. Hughes, and W. C. Sessa (1997)
J. Cell Biol. 137, 1525-1535
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Flotillin and Epidermal Surface Antigen Define a New Family of Caveolae-associated Integral Membrane Proteins.
P. E. Bickel, P. E. Scherer, J. E. Schnitzer, P. Oh, M. P. Lisanti, and H. F. Lodish (1997)
J. Biol. Chem. 272, 13793-13802
   Abstract »    Full Text »    PDF »
Murine SR-BI, a High Density Lipoprotein Receptor That Mediates Selective Lipid Uptake, Is N-Glycosylated and Fatty Acylated and Colocalizes with Plasma Membrane Caveolae.
J. Babitt, B. Trigatti, A. Rigotti, E. J. Smart, R. G. W. Anderson, S. Xu, and M. Krieger (1997)
J. Biol. Chem. 272, 13242-13249
   Abstract »    Full Text »    PDF »
Organized Endothelial Cell Surface Signal Transduction in Caveolae Distinct from Glycosylphosphatidylinositol-anchored Protein Microdomains.
J. Liu, P. Oh, T. Horner, R. A. Rogers, and J. E. Schnitzer (1997)
J. Biol. Chem. 272, 7211-7222
   Abstract »    Full Text »    PDF »
Loss of Caveolae, Vascular Dysfunction, and Pulmonary Defects in Caveolin-1 Gene-Disrupted Mice.
M. Drab, P. Verkade, M. Elger, M. Kasper, M. Lohn, B. Lauterbach, J. Menne, C. Lindschau, F. Mende, F. C. Luft, et al. (2001)
Science 293, 2449-2452
   Abstract »    Full Text »    PDF »
PC12 Cells Have Caveolae That Contain TrkA. CAVEOLAE-DISRUPTING DRUGS INHIBIT NERVE GROWTH FACTOR-INDUCED, BUT NOT EPIDERMAL GROWTH FACTOR-INDUCED, MAPK PHOSPHORYLATION.
S. Peiro, J. X. Comella, C. Enrich, D. Martin-Zanca, and N. Rocamora (2000)
J. Biol. Chem. 275, 37846-37852
   Abstract »    Full Text »    PDF »
Cholera Toxin Is Found in Detergent-insoluble Rafts/Domains at the Cell Surface of Hippocampal Neurons but Is Internalized via a Raft-independent Mechanism.
H. Shogomori and A. H. Futerman (2001)
J. Biol. Chem. 276, 9182-9188
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Replacement of the Transmembrane Anchor in Angiotensin I-converting Enzyme (ACE) with a Glycosylphosphatidylinositol Tail Affects Activation of the B2 Bradykinin Receptor by ACE Inhibitors.
B. Marcic, P. A. Deddish, R. A. Skidgel, E. G. Erdos, R. D. Minshall, and F. Tan (2000)
J. Biol. Chem. 275, 16110-16118
   Abstract »    Full Text »    PDF »
Distinction between signaling mechanisms in lipid rafts vs. caveolae.
G. Sowa, M. Pypaert, and W. C. Sessa (2001)
PNAS 98, 14072-14077
   Abstract »    Full Text »    PDF »


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