Acute cholesterol depletion inhibits clathrin-coated pit budding

Subtil, Agathe; Gaidarov, Ibragim; Kobylarz, Keith; Lampson, Michael A.; Keen, James H.; McGraw, Timothy E.

doi:10.1073/pnas.96.12.6775

Cited by 537 publications

(496 citation statements)

References 49 publications

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“…The finding that Tf-R and ASGP-R remained partially colocalized in treated cells further implies Tf-R delivery to recycling endosomes via early endosomes was maintained. Based on studies performed in cholesterol-depleted Chinese hamster ovary cells where Tf-R recycling to the PM was unchanged (Subtil et al, 1999), we further propose Tf-R recycled normally to the basolateral PM in lipid-depleted WIF-B cells.…”

Section: Cholesterol and Glycosphingolipids Are Specifically Requiredmentioning

confidence: 81%

Transcytotic Efflux from Early Endosomes Is Dependent on Cholesterol and Glycosphingolipids in Polarized Hepatic Cells

Nyasae

Hubbard

Tuma

2003

MBoC

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We examined the role that lipid rafts play in regulating apical protein trafficking in polarized hepatic cells. Rafts are postulated to form in the trans-Golgi network where they recruit newly synthesized apical residents and mediate their direct transport to the apical plasma membrane. In hepatocytes, single transmembrane and glycolipid-anchored apical proteins take the "indirect" route. They are transported from the trans-Golgi to the basolateral plasma membrane where they are endocytosed and transcytosed to the apical surface. Do rafts sort hepatic apical proteins along this circuitous pathway? We took two approaches to answer this question. First, we determined the detergent solubility of selected apical proteins and where in the biosynthetic pathway insolubility was acquired. Second, we used pharmacological agents to deplete raft components and assessed their effects on basolateral-to-apical transcytosis. We found that cholesterol and glycosphingolipids are required for delivery from basolateral early endosomes to the subapical compartment. In contrast, fluid phase uptake and clathrin-mediated internalization of recycling receptors were only mildly impaired. Apical protein solubility did not correlate with raft depletion or impaired transcytosis, suggesting other factors contribute to apical protein insolubility. Examination of apical proteins in Fao cells also revealed that raft-dependent sorting does not require the polarized cell context.

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Section: Cholesterol and Glycosphingolipids Are Specifically Requiredmentioning

confidence: 81%

Transcytotic Efflux from Early Endosomes Is Dependent on Cholesterol and Glycosphingolipids in Polarized Hepatic Cells

Nyasae

Hubbard

Tuma

2003

MBoC

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“…When COS-7 cells were transfected with dominant-negative dynamin (K44A), internalization and vesicle pinch-off by ST treatment were blocked (Figure 3c), indicating that, like EGF, ST-induced EGFR internalization required functional dynamin (Conner and Schmid, 2003). Cholesterol depletion also blocked EGFR internalization ( Figure 3d), suggesting that the cholesterol content played an important role in the internalization of EGFR induced by CA, similar to ligand-binding-induced EGFR internalization (Rodal et al, 1999;Subtil et al, 1999).…”

Section: Egfr Degradation Does Not Require Its Internalizationmentioning

confidence: 89%

Cleavage of epidermal growth factor receptor by caspase during apoptosis is independent of its internalization

Huang

2005

Oncogene

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“…When applied to cells, these cholesterol-binding drugs have effects other than their ability to extract cholesterol from membranes and disrupt L o domains. Perturbation of the actin cytoskeleton 59 and inhibition of clathrin-mediated endocytosis are probably directly related to cholesterol depletion per se [60][61][62] , but β-methyl-cyclodextrin has other effects that are unrelated to cholesterol depletion, such as global inhibition of the lateral mobility of plasma-membrane proteins, irrespective of their putative association with L o domains 63 . The latter effects are not seen with cholesterol depletion using statins 47,63,64 .…”

Section: Box 2 Limitations Of Biochemical Approaches To Study Lipid Rmentioning

confidence: 99%

Lipid rafts: contentious only from simplistic standpoints

Hancock

2006

Nat Rev Mol Cell Biol

742

716

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The hypothesis that lipid rafts exist in plasma membranes and have crucial biological functions remains controversial. The lateral heterogeneity of proteins in the plasma membrane is undisputed, but the contribution of cholesterol-dependent lipid assemblies to this complex, non-random organization promotes vigorous debate. In the light of recent studies with model membranes, computational modelling and innovative cell biology, I propose an updated model of lipid rafts that readily accommodates diverse views on plasma-membrane micro-organization.A widely accepted hypothesis in contemporary cell biology is that freely diffusing, stable, lateral assemblies of sphingolipids and cholesterol, which are termed lipid rafts 1-3 , constitute an important organizing principle for the plasma membrane. The basic concept is that lipid rafts can facilitate selective protein-protein interactions by selectively excluding or including proteins. This lipid-based sorting mechanism has been widely implicated in the assembly of transient signalling platforms and more permanent structures such as the immunological synapse, as well as in the sorting of proteins for entry into specific exocytic and endocytic trafficking pathways [1][2][3] . Despite the undoubted theoretical utility of lipid rafts to many cell biological processes, the basic hypothesis that stable lipid rafts exist at all in biological membranes is under intense scrutiny 4,5 . This is partly because lipid rafts, if they exist in resting cell membranes, are too small to be resolved by fluorescent microscopy and have no defined ultrastructure; therefore, proving their existence is problematic. This article will consider the biophysical properties of model membranes that underpin the lipid raft hypothesis, and the limitations of the biochemical approaches that have been used to study rafts in biological membranes that largely account for the current debate on the hypothesis mentioned above. After examining the challenges that are involved in correlating observations, which have been made in model and cellular membranes, I focus on synthesizing recent data on the size of lipid domains in model membranes with observations that have been obtained by imaging intact plasma membranes. These imaging approaches include single particle tracking (SPT), single fluorophore video tracking (SFVT), fluorescence resonance energy transfer (FRET), homo-FRET and electron microscopy (EM). On the one hand, these studies challenge the simplistic null hypothesis that lipid-based assemblies, such as lipid rafts, do not exist in biological membranes. On the other hand, it is timely to reconsider the raft hypothesis in the light of these new data because the consensus model that emerges is more complex than a simplistic notion of stable, freely diffusing lipid rafts. Some intriguing new questions aboutCompeting interests statement The author declares no competing financial interests. DATABASES

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Acute cholesterol depletion inhibits clathrin-coated pit budding

Cited by 537 publications

References 49 publications

Transcytotic Efflux from Early Endosomes Is Dependent on Cholesterol and Glycosphingolipids in Polarized Hepatic Cells

Transcytotic Efflux from Early Endosomes Is Dependent on Cholesterol and Glycosphingolipids in Polarized Hepatic Cells

Cleavage of epidermal growth factor receptor by caspase during apoptosis is independent of its internalization

Lipid rafts: contentious only from simplistic standpoints

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