title: Systems survey of endocytosis by multiparametric image analysis.
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.
authors.affiliation: Max Planck Institute for Molecular Cell Biology and Genetics, Germany.
authors.forename: Claudio
authors.initials: C
authors.lastname: Collinet
authors.forename: Martin
authors.initials: M
authors.lastname: Stöter
authors.forename: Charles R
authors.initials: CR
authors.lastname: Bradshaw
authors.forename: Nikolay
authors.initials: N
authors.lastname: Samusik
authors.forename: Jochen C
authors.initials: JC
authors.lastname: Rink
authors.forename: Denise
authors.initials: D
authors.lastname: Kenski
authors.forename: Bianca
authors.initials: B
authors.lastname: Habermann
authors.forename: Frank
authors.initials: F
authors.lastname: Buchholz
authors.forename: Robert
authors.initials: R
authors.lastname: Henschel
authors.forename: Matthias S
authors.initials: MS
authors.lastname: Mueller
authors.forename: Wolfgang E
authors.initials: WE
authors.lastname: Nagel
authors.forename: Eugenio
authors.initials: E
authors.lastname: Fava
authors.forename: Yannis
authors.initials: Y
authors.lastname: Kalaidzidis
authors.forename: Marino
authors.initials: M
authors.lastname: Zerial
mesh_terms: D003205:Computing Methodologies; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods*; D055106:Genome-Wide Association Study; D006801:Humans; D007091:Image Processing, Computer-Assisted; D053858:Metabolic Networks and Pathways / Q000502:physiology; D018613:Microscopy, Confocal; D010641:Phenotype; D021381:Protein Transport / Q000502:physiology; D034622:RNA Interference; D015398:Signal Transduction / Q000502:physiology; D014168:Transferrin / Q000378:metabolism
chemical_list: D014168:Transferrin; D004815:Epidermal Growth Factor