Spatial control of coated-pit dynamics in living cells

Gaidarov, Ibragim; Santini, Francesca; Warren, Robin A.; Keen, James H.

doi:10.1038/8971

Cited by 396 publications

(453 citation statements)

References 46 publications

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“…Functional and morphological control experiments are necessary to remove such doubt. Again, using clathrin-coated pits as an example, we and others have shown that when FP-tagged clathrin-light chain A (CLa) was expressed in cells in the presence of endogenous CLa, more than 96% of clathrin-coated pits visualized by immunofluorescence or with fluorescently labelled transferrin co-localize with FP-marked structures 13,14 , showing that nearly all clathrincoated pits are detected by the FP signal.…”

Section: Virus and Cell Labelingmentioning

confidence: 92%

Virus trafficking – learning from single-virus tracking

2007

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What could be a better way to study virus trafficking than 'miniaturizing oneself' and 'taking a ride with the virus particle' on its journey into the cell? Single-virus tracking in living cells potentially provides us with the means to visualize the virus journey. This approach allows us to follow the fate of individual virus particles and monitor dynamic interactions between viruses and cellular structures, revealing previously unobservable infection steps. The entry, trafficking and egress mechanisms of various animal viruses have been elucidated using this method. The combination of single-virus trafficking with systems approaches and state-of-the-art imaging technologies should prove exciting in the future.

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Section: Virus and Cell Labelingmentioning

confidence: 92%

Virus trafficking – learning from single-virus tracking

2007

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“…Involvement of the actin cortex in the mobility of GFP-Iabelled clathrin-coated pits was recently shown in living mammalian (Gaidarov et al 1999) and D. discoideum cells (Damer et al 1998; Mol Bioi Cell 9: 130a).…”

Section: Molecular Aspects Of the Internalisation Processesmentioning

confidence: 92%

Molecular Mechanisms of Membrane Trafficking. What do we Learn from Dictyostelium discoideum?

Neuhaus

Soldati

1999

Protist

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“…In contrast, label for Thy-1 is neither concentrated in nor excluded from the patches or their neighboring coated pits, suggesting a relative absence of topographical constraints for this GPI-linked protein even after external cross-linking. It is known that clathrin-coated pits bud from topographically restricted membrane regions, including the uropods of polarized macrophages and neutrophils and the cleavage furrows of dividing cells (Pfeiffer et al, 1980;Oliver and Berlin, 1983) and that multiple vesicles may form from a single pit (Gaidarov et al, 1999;illustrated in Wilson et al, 2000). This, and other recent work (Blanpied et al, 2002), suggests cells can organize "hot spots" that are sites of repeated rounds of clathrin vesicle formation.…”

Section: Discussionmentioning

confidence: 99%

Markers for Detergent-resistant Lipid Rafts Occupy Distinct and Dynamic Domains in Native Membranes

Wilson

Steinberg

Liederman

et al. 2004

MBoC

187

183

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Lipid rafts isolated by detergent extraction and sucrose gradient fractionation from mast cells are enriched for the glycosylphosphatidylinositol-linked protein Thy-1, the ganglioside GM1, palmitoylated LAT, and cross-linked IgE receptors, Fc⑀RI. This study addresses the relationship of fractionation data to the organization of raft markers in native membranes. Immunogold labeling and electron microscopy shows there is little or no colocalization of the raft markers Thy-1, GM1, and LAT with each other or with Fc⑀RI on native membrane sheets prepared from unstimulated cells. External cross-linking of Thy-1 promotes coclustering of Thy-1 with LAT, but not with GM1. Thy-1 and LAT clusters occur on membrane regions without distinctive features. In contrast, external cross-linking of Fc⑀RI and GM1 causes their redistribution to electron-dense membrane patches independently of each other and of Thy-1. The distinctive patches that accumulate cross-linked Fc⑀RI and GM1 also accumulate osmium, a stain for unsaturated lipids, and are sites for coated vesicle budding. Electron microscopy reveals a more complex and dynamic topographical organization of membrane microdomains than is predicted by biochemical analysis of detergent-resistant membranes. INTRODUCTIONOrdered regions of membrane, known as microdomains, lipid rafts, detergent-resistant membranes (DRMs), and other abbreviations are thought to be critical sites of signal propagation and membrane trafficking (Edidin, 1997(Edidin, , 2001Simons and Ikonen, 1997;Anderson, 1998; London, 1998, 2000;Jacobson and Dietrich, 1999;Langlet et al., 2000;Anderson and Jacobson, 2002). Analysis of these regions typically begins with detergent solubilization of whole cells followed by sucrose density gradient centrifugation and the recovery of detergent-resistant membranes from the light fractions of the gradient. The DRMs are enriched for caveolin, glycosylphosphatidylinositol-linked (GPI-linked) proteins, glycosphingolipids, GM1 ganglioside, and cholesterol, suggesting that these components are associated in the liquid-ordered (l o ) phase of the lipid bilayer (Schroeder et al., 1994;Ahmed et al., 1997). The interpretation of gradient centrifugation experiments remains controversial. There is evidence that detergents may force associations between components that are not colocalized in intact cells (Mayor and Maxfield, 1995), and fractionation results are known to be dramatically altered by varying the concentration of Triton X-100 (Field et al., 1999;Parolini et al., 1999), by use of alternative detergents (Montixi et al., 1998;Surviladze et al., 1998) or by omission of detergent altogether (Ilangumaran et al., 1999;Harder and Kuhn, 2000;Surviladze et al., 2001). Methods to observe membrane segregation in situ have also generated controversy. Results based on light and fluorescence microscopy of proteins and lipids, including refinements of the established fluorescence recovery after photobleaching methods and new single particle tracking methods, have led some investigators to co...

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Spatial control of coated-pit dynamics in living cells

Cited by 396 publications

References 46 publications

Virus trafficking – learning from single-virus tracking

Virus trafficking – learning from single-virus tracking

Molecular Mechanisms of Membrane Trafficking. What do we Learn from Dictyostelium discoideum?

Markers for Detergent-resistant Lipid Rafts Occupy Distinct and Dynamic Domains in Native Membranes

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