Corrigendum: Systems survey of endocytosis by multiparametric image analysis

Max Planck Institute for Molecular Cell Biology and Genetics, Germany.
Nature (Impact Factor: 41.46). 02/2010; 464(7286):243-9. DOI: 10.1038/nature08779
Source: PubMed

ABSTRACT Endocytosis is a complex process fulfilling many cellular and developmental functions. Understanding how it is regulated and integrated with other cellular processes requires a comprehensive analysis of its molecular constituents and general design principles. Here, we developed a new strategy to phenotypically profile the human genome with respect to transferrin (TF) and epidermal growth factor (EGF) endocytosis by combining RNA interference, automated high-resolution confocal microscopy, quantitative multiparametric image analysis and high-performance computing. We identified several novel components of endocytic trafficking, including genes implicated in human diseases. We found that signalling pathways such as Wnt, integrin/cell adhesion, transforming growth factor (TGF)-beta and Notch regulate the endocytic system, and identified new genes involved in cargo sorting to a subset of signalling endosomes. A systems analysis by Bayesian networks further showed that the number, size, concentration of cargo and intracellular position of endosomes are not determined randomly but are subject to specific regulation, thus uncovering novel properties of the endocytic system.

23 Reads
  • Source
    • "Interestingly, a functional genomics screen revealed that the endosomal system is modulated by a number of metabolic pathways, including glycolysis, gluconeogenesis, and steroid biosynthesis (Collinet et al., 2010). This argues that the functional relationship between metabolism and endocytosis is bi-directional . "
    [Show abstract] [Hide abstract]
    ABSTRACT: The liver maintains glucose and lipid homeostasis by adapting its metabolic activity to the energy needs of the organism. Communication between hepatocytes and extracellular environment via endocytosis is key to such homeostasis. Here, we addressed the question of whether endosomes are required for gluconeogenic gene expression. We took advantage of the loss of endosomes in the mouse liver upon Rab5 silencing. Strikingly, we found hepatomegaly and severe metabolic defects such as hypoglycemia, hypercholesterolemia, hyperlipidemia, and glycogen accumulation that phenocopied those found in von Gierke's disease, a glucose-6-phosphatase (G6Pase) deficiency. G6Pase deficiency alone can account for the reduction in hepatic glucose output and glycogen accumulation as determined by mathematical modeling. Interestingly, we uncovered functional alterations in the transcription factors, which regulate G6Pase expression. Our data highlight a requirement of Rab5 and the endosomal system for the regulation of gluconeogenic gene expression that has important implications for metabolic diseases. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.
    Cell Reports 04/2015; 11(6). DOI:10.1016/j.celrep.2015.04.018 · 8.36 Impact Factor
  • Source
    • "However, we did not find any significant effect of PLCG1 knockdown on the number of transferrin receptor–containing endosomes (Supplemental Figure S6A). The number of endosomes has been used to infer effects on the endocytic system (Collinet et al., 2010). In addition, searching the endosomics database (endosomics "
    [Show abstract] [Hide abstract]
    ABSTRACT: The role of early secretory trafficking in the regulation of cell motility remains incompletely understood. Here, we used a siRNA screen monitoring effects on structure of the Golgi apparatus and on cell migration. Two major Golgi phenotypes were observed, fragmented and small Golgi. The latter exhibited a stronger correlation with a defect in cell migration. Among the small Golgi hits, we focused on phospholipase C gamma1 (PLCγ1). We show that PLCγ1 regulates Golgi structure and cell migration independently from its catalytic activity, but in a manner dependent on interaction with the tethering protein p115. PLCγ1 regulates the dynamics of p115 in the early secretory pathway, thereby controling trafficking from the endoplasmic reticulum to the Golgi. Our results uncover a new function of PLCγ1 that is independent of its catalytic function and link early secretory trafficking to the regulation of cell migration. © 2015 by The American Society for Cell Biology.
    Molecular biology of the cell 04/2015; 26(12). DOI:10.1091/mbc.E15-03-0178 · 4.47 Impact Factor
  • Source
    • "Cell–cell communication is essential for the development and homeostasis of multicellular organisms. Signaling by cell surface receptors and endocytosis are highly integrated processes (Seto et al., 2002; Le Borgne et al., 2005; Polo and Di Fiore, 2006; Collinet et al., 2010). The interplay between membrane trafficking and cell–cell signaling has been well studied in the context of sensory organs in Drosophila melanogaster (Kandachar and Roegiers, 2012). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Signaling and endocytosis are highly integrated processes that regulate cell fate. In the Drosophila melanogaster sensory bristle lineages, Numb inhibits the recycling of Notch and its trafficking partner Sanpodo (Spdo) to regulate cell fate after asymmetric cell division. In this paper, we have used a dual GFP/Cherry tagging approach to study the distribution and endosomal sorting of Notch and Spdo in living pupae. The specific properties of GFP, i.e., quenching at low pH, and Cherry, i.e., slow maturation time, revealed distinct pools of Notch and Spdo: cargoes exhibiting high GFP/low Cherry fluorescence intensities localized mostly at the plasma membrane and early/sorting endosomes, whereas low GFP/high Cherry cargoes accumulated in late acidic endosomes. These properties were used to show that Spdo is sorted toward late endosomes in a Numb-dependent manner. This dual-tagging approach should be generally applicable to study the trafficking dynamics of membrane proteins in living cells and tissues.
    The Journal of Cell Biology 11/2014; 207(3). DOI:10.1083/jcb.201407071 · 9.83 Impact Factor
Show more