Endosome maturation. EMBO J

Institute of Biochemistry, ETH Zurich, Switzerland.
The EMBO Journal (Impact Factor: 10.43). 08/2011; 30(17):3481-500. DOI: 10.1038/emboj.2011.286
Source: PubMed


Being deeply connected to signalling, cell dynamics, growth, regulation, and defence, endocytic processes are linked to almost all aspects of cell life and disease. In this review, we focus on endosomes in the classical endocytic pathway, and on the programme of changes that lead to the formation and maturation of late endosomes/multivesicular bodies. The maturation programme entails a dramatic transformation of these dynamic organelles disconnecting them functionally and spatially from early endosomes and preparing them for their unidirectional role as a feeder pathway to lysosomes.

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Available from: Ari Helenius, Jan 13, 2014
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    • "Direct immunofluorescence studies and co-labeling with Lysotracker Red, a marker for acidic late endosomes and lysosomes, further showed that the internalized antibody was trafficked to acidic organelles in the perinuclear cytoplasm. The known perinuclear localization of lysosomes and late endosomes (Huotari and Helenius, 2011) agreed with the localization of OX42-fluorescence seen here confirming lysosomal trafficking of OX42 antibody in microglia. Lysosomal trafficking is desirable for non-viral vehicles that utilize PEI as DNA condensation agent, because the gradual acidification of endosomes facilitates the endosomal escape of the gene vehicle (Varkouhi et al., 2011). "
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    ABSTRACT: Microglial activation is a central event in neurodegeneration. Novel technologies are sought for that specifically manipulate microglial function in order to delineate their role in onset and progression of neuropathologies. We investigated for the first time whether non-viral gene delivery based on polyethyleneglycol (PEG)-polyethyleneimine (PEI) conjugated to the monoclonal anti-CD11b antibody OX42 (‘OX42-immunogene’) could be used to specifically target microglia. We identified the microglial integrin receptor CD11b as a potential target for receptor-mediated gene transfer based on its cellular specificity in mixed glia culture and in vivo and found that the OX42 antibody is rapidly internalized and trafficked to acidic organelles in absence of activation of the respiratory burst. We then showed that the OX42-immunogene although internalised was degraded intracellularly and did not cause substantial gene expression in microglia. Investigation of specific barriers to microglial gene transfer revealed that aggregated OX42-immunogene polyplexes stimulated the respiratory burst that likely involved Fcγ-receptors. Transfections in the presence of the endosomolytic agent chloroquine improved transfection efficiency indicating that endosomal escape may be limited. This study identifies CD11b as an entry point for antibody-mediated gene transfer into microglia and takes important steps towards the further development of OX42-immunogenes.
    Frontiers in Molecular Neuroscience 10/2014; 7. DOI:10.3389/fnmol.2014.00079 · 4.08 Impact Factor
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    • "Thus degradation of the desmosomal proteins other than DP was blocked by lysosomal inhibitors and the internalised material co-localised with the lysosomal marker Lamp1. Lysosomal degradation was suggested previously by association of half desmosomes with late endosomes [46], structures which fuse with lysosomes [70]. "
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    ABSTRACT: Desmosomes are intercellular adhesive junctions of major importance for tissue integrity. To allow cell motility and migration they are down-regulated in epidermal wound healing. Electron microscopy indicates that whole desmosomes are internalised by cells in tissues, but the mechanism of down-regulation is unclear. In this paper we provide an overview of the internalisation of half-desmosomes by cultured cells induced by calcium chelation. Our results show that: (i) half desmosome internalisation is dependent on conventional PKC isoforms; (ii) microtubules transport internalised half desmosomes to the region of the centrosome by a kinesin-dependent mechanism; (iii) desmosomal proteins remain colocalised after internalisation and are not recycled to the cell surface; (iv) internalised desmosomes are degraded by the combined action of lysosomes and proteasomes. We also confirm that half desmosome internalisation is dependent upon the actin cytoskeleton. These results suggest that half desmosomes are not disassembled and recycled during or after internalisation but instead are transported to the centrosomal region where they are degraded. These findings may have significance for the down-regulation of desmosomes in wounds.
    PLoS ONE 10/2014; 9(10):e108570. DOI:10.1371/journal.pone.0108570 · 3.23 Impact Factor
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    • "Signal transduction by membrane receptors, protein delivery to specific compartments, membrane repair and exosome shedding or autophagy, among others, depend on compartmentalized traffic of molecules within the cell. Intracellular vesicles constitute a yet not fully characterized array of dynamic membranous compartments which are in continuous evolution and transformation, resulting in the acquisition of a specific protein and lipid composition [1], [2]. These components can regulate the fate of proteins by sorting them through different types of intracellular vesicles that can come from or recycle back to the plasma membrane, send proteins along the secretory pathway from the Golgi or travel along trafficking routes through the late endosomal pathway to the lysosome to be degraded, among many other processes. "
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    ABSTRACT: The C-terminal ends of small GTPases contain hypervariable sequences which may be posttranslationally modified by defined lipid moieties. The diverse structural motifs generated direct proteins towards specific cellular membranes or organelles. However, knowledge on the factors that determine these selective associations is limited. Here we show, using advanced microscopy, that the isoprenylation and palmitoylation motif of human RhoB (-CINCCKVL) targets chimeric proteins to intraluminal vesicles of endolysosomes in human cells, displaying preferential co-localization with components of the late endocytic pathway. Moreover, this distribution is conserved in distant species, including cells from amphibians, insects and fungi. Blocking lipidic modifications results in accumulation of CINCCKVL chimeras in the cytosol, from where they can reach endolysosomes upon release of this block. Remarkably, CINCCKVL constructs are sorted to intraluminal vesicles in a cholesterol-dependent process. In the lower species, neither the C-terminal sequence of RhoB, nor the endosomal distribution of its homologs are conserved; in spite of this, CINCCKVL constructs also reach endolysosomes in Xenopus laevis and insect cells. Strikingly, this behavior is prominent in the filamentous ascomycete fungus Aspergillus nidulans, in which GFP-CINCCKVL is sorted into endosomes and vacuoles in a lipidation-dependent manner and allows monitoring endosomal movement in live fungi. In summary, the isoprenylated and palmitoylated CINCCKVL sequence constitutes a specific structure which delineates an endolysosomal sorting strategy operative in phylogenetically diverse organisms.
    PLoS ONE 09/2014; 9(9):e107190. DOI:10.1371/journal.pone.0107190 · 3.23 Impact Factor
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