Related Experiment Video
Updated: Apr 23, 2026
08:49
Cell Surface Receptor Identification Using Genome-Scale CRISPR/Cas9 Genetic Screens
Published on: June 6, 2020
15.2K
A high-throughput siRNA screen identifies genes that regulate mannose 6-phosphate receptor trafficking
Mihaela Anitei1, Ramu Chenna1, Cornelia Czupalla1
1Biotechnology Center, Technische Universität Dresden, Tatzberg 47-51, 01307 Dresden, Germany.
Journal of Cell Science
|October 4, 2014
Summary
Cell signaling regulates the trafficking of mannose 6-phosphate receptors (MPRs) crucial for lysosome function. Researchers identified key kinases and phosphatases involved in MPR pathways, revealing a SRC-dependent mechanism controlling carrier biogenesis.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Lysosomal hydrolase delivery to lysosomes is vital for cellular homeostasis.
- Mannose 6-phosphate receptors (MPRs) mediate this delivery via specific trafficking pathways.
- Dysregulated MPR trafficking impacts various cellular processes.
Purpose of the Study:
- To investigate how cell signaling pathways influence MPR trafficking.
- To identify kinases and phosphatases regulating MPR dynamics and associated subcellular compartments.
- To elucidate the signaling mechanisms controlling post-Golgi carrier biogenesis.
Main Methods:
- High-throughput RNA interference (RNAi) screening of the human kinome and phosphatome.
- High-content image analysis to assess MPR trafficking and compartment dynamics.
- Analysis of phosphatidylinositol phosphate metabolism enzymes in MPR pathways.
Main Results:
- Identified 127 kinases and phosphatases impacting MPR trafficking and cellular compartment dynamics.
- Mapped MPR trafficking pathways involving enzymes that regulate phosphatidylinositol phosphate metabolism.
- Discovered a SRC-dependent signaling pathway controlling ARF1, RAC1, and myosin II activity in post-Golgi carrier biogenesis.
Conclusions:
- Cell signaling critically modulates MPR trafficking pathways.
- Phosphatidylinositol phosphate metabolism is a key regulatory node in MPR transport.
- SRC signaling orchestrates the formation of endosomal carriers via ARF1/RAC1 and myosin II.
