![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 17, Issue 7, 3197-3210, July 2006
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
*Department of Biochemistry and Molecular Biology, Thoracic Diseases Research Unit, Mayo Clinic College of Medicine, Rochester, MN 55905; and Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo 162-8640, Japan
Submitted December 5, 2005;
Accepted April 24, 2006
Sphingolipids (SLs) play important roles in membrane structure and cell function. Here, we examine the SL requirements of various endocytic mechanisms using a mutant cell line and pharmacological inhibitors to disrupt SL biosynthesis. First, we demonstrated that in Chinese hamster ovary cells we could distinguish three distinct mechanisms of clathrin-independent endocytosis (caveolar, RhoA, and Cdc42 dependent) which differed in cargo, sensitivity to pharmacological agents, and dominant negative proteins. General depletion of SLs inhibited endocytosis by each clathrin-independent mechanism, whereas clathrin-dependent uptake was unaffected. Depletion of glycosphingolipids (GSLs; a subgroup of SLs) selectively blocked caveolar endocytosis and decreased caveolin-1 and caveolae at the plasma membrane. Caveolar endocytosis and PM caveolae could be restored in GSL-depleted cells by acute addition of exogenous GSLs. Disruption of RhoA- and Cdc42-regulated endocytosis by SL depletion was shown to be related to decreased targeting of these Rho proteins to the plasma membrane and could be partially restored by exogenous sphingomyelin but not GSLs. Both the in vivo membrane targeting and in vitro binding to artificial lipid vesicles of RhoA and Cdc42 were shown to be dependent upon sphingomyelin. These results provide the first evidence that SLs are differentially required for distinct mechanisms of clathrin-independent endocytosis.
The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).
Present address: Photometrics, 3440 East Britannia Dr., Tucson, AZ 85706.
Address correspondence to: Richard E. Pagano ( pagano.richard{at}mayo.edu)
This article has been cited by other articles:
|
|
 |
|
  D. J. Barr, A. G. Ostermeyer-Fay, R. A. Matundan, and D. A. Brown Clathrin-independent endocytosis of ErbB2 in geldanamycin-treated human breast cancer cells J. Cell Sci., October 1, 2008; 121(19): 3155 - 3166. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Klima and I. Foissner FM Dyes Label Sterol-Rich Plasma Membrane Domains and are Internalized Independently of the Cytoskeleton in Characean Internodal Cells Plant Cell Physiol., October 1, 2008; 49(10): 1508 - 1521. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Galvan, P. G. Camoletto, F. Cristofani, P. P. Van Veldhoven, and M. D. Ledesma Anomalous Surface Distribution of Glycosyl Phosphatidyl Inositol-anchored Proteins in Neurons Lacking Acid Sphingomyelinase Mol. Biol. Cell, February 1, 2008; 19(2): 509 - 522. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Yamauchi, P. C. Reid, J. B. Sperry, K. Furukawa, M. Takeya, C. C. Y. Chang, and T.-Y. Chang Plasma Membrane Rafts Complete Cholesterol Synthesis by Participating in Retrograde Movement of Precursor Sterols J. Biol. Chem., November 30, 2007; 282(48): 34994 - 35004. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Nishi and K. Saigo Cellular Internalization of Green Fluorescent Protein Fused with Herpes Simplex Virus Protein VP22 via a Lipid Raft-mediated Endocytic Pathway Independent of Caveolae and Rho Family GTPases but Dependent on Dynamin and Arf6 J. Biol. Chem., September 14, 2007; 282(37): 27503 - 27517. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Tateno, H. Li, M. J. Schur, N. Bovin, P. R. Crocker, W. W. Wakarchuk, and J. C. Paulson Distinct Endocytic Mechanisms of CD22 (Siglec-2) and Siglec-F Reflect Roles in Cell Signaling and Innate Immunity Mol. Cell. Biol., August 15, 2007; 27(16): 5699 - 5710. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. C. Dorsey, T. Muthusamy, M. A. Whitt, and J. V. Cox A novel role for a YXX{Phi} motif in directing the caveolin-dependent sorting of membrane-spanning proteins J. Cell Sci., August 1, 2007; 120(15): 2544 - 2554. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. C. Gauthier, P. Monzo, T. Gonzalez, A. Doye, A. Oldani, P. Gounon, V. Ricci, M. Cormont, and P. Boquet Early endosomes associated with dynamic F-actin structures are required for late trafficking of H. pylori VacA toxin J. Cell Biol., April 23, 2007; 177(2): 343 - 354. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Kippert, K. Trajkovic, L. Rajendran, J. Ries, and M. Simons Rho Regulates Membrane Transport in the Endocytic Pathway to Control Plasma Membrane Specialization in Oligodendroglial Cells J. Neurosci., March 28, 2007; 27(13): 3560 - 3570. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. D. Singh, E. L. Holicky, Z.-j. Cheng, S.-Y. Kim, C. L. Wheatley, D. L. Marks, R. Bittman, and R. E. Pagano Inhibition of caveolar uptake, SV40 infection, and {beta}1-integrin signaling by a nonnatural glycosphingolipid stereoisomer J. Cell Biol., March 26, 2007; 176(7): 895 - 901. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. D. Singh, Y. Liu, C. L. Wheatley, E. L. Holicky, A. Makino, D. L. Marks, T. Kobayashi, G. Subramaniam, R. Bittman, and R. E. Pagano Caveolar Endocytosis and Microdomain Association of a Glycosphingolipid Analog Is Dependent on Its Sphingosine Stereochemistry J. Biol. Chem., October 13, 2006; 281(41): 30660 - 30668. [Abstract] [Full Text] [PDF]
|
|
