Systems survey of endocytosis by multiparametric image analysis.
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.
Collinet
Claudio
C
Max Planck Institute for Molecular Cell Biology and Genetics, Germany.
Stöter
Martin
M
Bradshaw
Charles R
CR
Samusik
Nikolay
N
Rink
Jochen C
JC
Kenski
Denise
D
Habermann
Bianca
B
Buchholz
Frank
F
Henschel
Robert
R
Mueller
Matthias S
MS
Nagel
Wolfgang E
WE
Fava
Eugenio
E
Kalaidzidis
Yannis
Y
Zerial
Marino
M
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
D014168:Transferrin; D004815:Epidermal Growth Factor