title: A high-throughput siRNA screen identifies genes that regulate mannose 6-phosphate receptor trafficking.
abstract: The delivery of newly synthesized soluble lysosomal hydrolases to the endosomal system is essential for lysosome function and cell homeostasis. This process relies on the proper trafficking of the mannose 6-phosphate receptors (MPRs) between the trans-Golgi network (TGN), endosomes and the plasma membrane. Many transmembrane proteins regulating diverse biological processes ranging from virus production to the development of multicellular organisms also use these pathways. To explore how cell signaling modulates MPR trafficking, we used high-throughput RNA interference (RNAi) to target the human kinome and phosphatome. Using high-content image analysis, we identified 127 kinases and phosphatases belonging to different signaling networks that regulate MPR trafficking and/or the dynamic states of the subcellular compartments encountered by the MPRs. Our analysis maps the MPR trafficking pathways based on enzymes regulating phosphatidylinositol phosphate metabolism. Furthermore, it reveals how cell signaling controls the biogenesis of post-Golgi tubular carriers destined to enter the endosomal system through a SRC-dependent pathway regulating ARF1 and RAC1 signaling and myosin II activity.
authors.affiliation: Biotechnology Center, Technische Universität Dresden, Tatzberg 47-51, 01307 Dresden, Germany.
authors.forename: Mihaela
authors.initials: M
authors.lastname: Anitei
authors.affiliation: Biotechnology Center, Technische Universität Dresden, Tatzberg 47-51, 01307 Dresden, Germany.
authors.forename: Ramu
authors.initials: R
authors.lastname: Chenna
authors.affiliation: Biotechnology Center, Technische Universität Dresden, Tatzberg 47-51, 01307 Dresden, Germany.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Czupalla
authors.affiliation: Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Kamenice 3, Building A1, 62500 Brno, Czech Republic.
authors.forename: Milan
authors.initials: M
authors.lastname: Esner
authors.affiliation: Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany.
authors.forename: Sara
authors.initials: S
authors.lastname: Christ
authors.affiliation: Biotechnology Center, Technische Universität Dresden, Tatzberg 47-51, 01307 Dresden, Germany.
authors.forename: Steffi
authors.initials: S
authors.lastname: Lenhard
authors.affiliation: Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Korn
authors.affiliation: Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany.
authors.forename: Felix
authors.initials: F
authors.lastname: Meyenhofer
authors.affiliation: Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany.
authors.forename: Marc
authors.initials: M
authors.lastname: Bickle
authors.affiliation: Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany.
authors.forename: Marino
authors.initials: M
authors.lastname: Zerial
authors.affiliation: Biotechnology Center, Technische Universität Dresden, Tatzberg 47-51, 01307 Dresden, Germany bernard.hoflack@biotec.tu-dresden.de.
authors.forename: Bernard
authors.initials: B
authors.lastname: Hoflack
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology*; D016000:Cluster Analysis; D011992:Endosomes / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic; D053263:Gene Regulatory Networks; D006367:HeLa Cells; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D060066:Protein Interaction Maps; D021381:Protein Transport / Q000235:genetics; D034622:RNA Interference; D017527:Receptor, IGF Type 2 / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection; D020830:rac1 GTP-Binding Protein / Q000235:genetics / Q000378:metabolism; D019061:src-Family Kinases / Q000235:genetics / Q000378:metabolism; D021601:trans-Golgi Network / Q000201:enzymology*
chemical_list: D018129:Phosphatidylinositol Phosphates; C107953:RAC1 protein, human; D017527:Receptor, IGF Type 2; D019061:src-Family Kinases; D020823:ADP-Ribosylation Factor 1; D020830:rac1 GTP-Binding Protein
keywords: Lysosome; Mannose 6-phosphate receptor; Post-Golgi transport; Trafficking; siRNA screen