![[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. 15, Issue 10, 4500-4511, October 2004
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Department of Biochemistry and Molecular Biology, Mayo Clinic and Foundation, Rochester, MN 55905
Submitted May 25, 2004;
Revised July 22, 2004;
Accepted July 23, 2004
Monitoring Editor: Suzanne Pfeffer
In normal human skin fibroblasts (HSFs), fluorescent glycosphingolipid analogues are endocytosed and sorted into two pools, one that is recycled to the plasma membrane and one that is transported to the Golgi complex. Here, we investigated glycosphingolipid recycling in Niemann-Pick type A and C lipid storage disease fibroblasts (NPFs). Cells were incubated with a fluorescent analogue of lactosylceramide (LacCer) at 16°C to label early endosomes (EEs), shifted to 37°C, and lipid recycling was quantified. Using dominant negative rabs, we showed that, in normal HSFs, LacCer recycling was rapid (t1/2 
8 min) and mainly rab4-dependent. In NPFs, LacCer recycling was delayed (t1/2 30–40 min), and rab4-dependent recycling was absent, whereas rab11-dependent recycling predominated. Transferrin recycling via the rab4 pathway was similarly perturbed in NPFs. Compared with normal HSFs, EEs in NPFs showed high cholesterol levels and an altered organization of rab4. In vitro extraction of rab4 (but not rab11) with GDP dissociation inhibitor was severely attenuated in NPF endosomal fractions. This impairment was reversed with cholesterol depletion of isolated endosomes or with high-salt treatment of endosomes. These data suggest that abnormal membrane recycling in NPFs results from specific inhibition of rab4 function by excess cholesterol in EEs.
Abbreviations used: BODIPY-LacCer, N-[5-(5,7-dimethylborondipyrromethenedifluoride)-1-penanoyl]-lactosylsphingosine; DF-BSA, defatted bovine serum albumin; DN, dominant negative; EE, early endosome; FITC, fluorescein isothiocyanate; GDI, GDP dissociation inhibitor; GFP, gene fluorescent protein; GSL, glycosphingolipid; GST, glutathione S-transferase; HSF, human skin fibroblast; LacCer, lactosylceramide; m
-CD, methyl-B-cyclodextrin; NP-A, Niemann-Pick disease, type A; NP-C, Niemann-Pick disease, type C; NPF, Niemann-Pick fibroblast; P13K, phosphoinositide 3-kinase; PM, plasma membrane; RE, recycling endosome; SL, sphingolipid; SLSD, sphingolipid storage disease; Tfn, transferrin; WM, wortmannin; WT, wild-type.
The online version of this article contains supplemental material accessible through http://www.molbiolcell.org.
* These authors contributed equally to this work.
Corresponding author. E-mail address: pagano.richard{at}mayo.edu.
This article has been cited by other articles:
|
|
 |
|
  H. Chen, J. Yang, P. S. Low, and J.-X. Cheng Cholesterol Level Regulates Endosome Motility via Rab Proteins Biophys. J., February 15, 2008; 94(4): 1508 - 1520. [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]
|
|
|
|
|
|
M. Koivusalo, M. Jansen, P. Somerharju, and E. Ikonen Endocytic Trafficking of Sphingomyelin Depends on Its Acyl Chain Length Mol. Biol. Cell, December 1, 2007; 18(12): 5113 - 5123. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Takahashi, M. Murate, M. Fukuda, S. B. Sato, A. Ohta, and T. Kobayashi Cholesterol Controls Lipid Endocytosis through Rab11 Mol. Biol. Cell, July 1, 2007; 18(7): 2667 - 2677. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. F. Moore, O. V. Krokhin, R. C. Beavis, M. Ries, C. Robinson, E. Goldin, R. O. Brady, J. A. Wilkins, and R. Schiffmann Proteomics of specific treatment-related alterations in Fabry disease: A strategy to identify biological abnormalities PNAS, February 20, 2007; 104(8): 2873 - 2878. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Gentzsch, A. Choudhury, X.-b. Chang, R. E. Pagano, and J. R. Riordan Misassembled mutant {Delta}F508 CFTR in the distal secretory pathway alters cellular lipid trafficking J. Cell Sci., February 1, 2007; 120(3): 447 - 455. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. D. Linder, R.-L. Uronen, M. Holtta-Vuori, P. van der Sluijs, J. Peranen, and E. Ikonen Rab8-dependent Recycling Promotes Endosomal Cholesterol Removal in Normal and Sphingolipidosis Cells Mol. Biol. Cell, January 1, 2007; 18(1): 47 - 56. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. G. Ng and D. E. Griffin Acid Sphingomyelinase Deficiency Increases Susceptibility to Fatal Alphavirus Encephalomyelitis J. Virol., November 15, 2006; 80(22): 10989 - 10999. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Sugii, S. Lin, N. Ohgami, M. Ohashi, C. C. Y. Chang, and T.-Y. Chang Roles of Endogenously Synthesized Sterols in the Endocytic Pathway J. Biol. Chem., August 11, 2006; 281(32): 23191 - 23206. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. G. Ganley and S. R. Pfeffer Cholesterol Accumulation Sequesters Rab9 and Disrupts Late Endosome Function in NPC1-deficient Cells J. Biol. Chem., June 30, 2006; 281(26): 17890 - 17899. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. H. Pipalia, A. Huang, H. Ralph, M. Rujoi, and F. R. Maxfield Automated microscopy screening for compounds that partially revert cholesterol accumulation in Niemann-Pick C cells J. Lipid Res., February 1, 2006; 47(2): 284 - 301. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. McCampbell, D. Truong, D. C. Broom, A. Allchorne, K. Gable, R. G. Cutler, M. P. Mattson, C. J. Woolf, M. P. Frosch, J. M. Harmon, et al. Mutant SPTLC1 dominantly inhibits serine palmitoyltransferase activity in vivo and confers an age-dependent neuropathy Hum. Mol. Genet., November 15, 2005; 14(22): 3507 - 3521. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T.-Y. Chang, P. C. Reid, S. Sugii, N. Ohgami, J. C. Cruz, and C. C. Y. Chang Niemann-Pick Type C Disease and Intracellular Cholesterol Trafficking J. Biol. Chem., June 3, 2005; 280(22): 20917 - 20920. [Full Text] [PDF]
|
|
