Abstract
Endocytosis mediates the cellular uptake of micronutrients and the turnover of plasma membrane proteins. Clathrin-mediated endocytosis is the major uptake pathway in resting cells1, but several clathrin-independent endocytic routes exist in parallel2,3. One such pathway, fast endophilin-mediated endocytosis (FEME), is not constitutive but triggered upon activation of certain receptors, including the β1 adrenergic receptor4. FEME activates promptly following stimulation as endophilin is pre-enriched by the phosphatidylinositol-3,4-bisphosphate-binding protein lamellipodin4,5. However, in the absence of stimulation, endophilin foci abort and disassemble after a few seconds. Looking for additional proteins involved in FEME, we found that 20 out of 65 BAR domain-containing proteins tested colocalized with endophilin spots. Among them, FBP17 and CIP4 prime the membrane of resting cells for FEME by recruiting the 5'-lipid phosphatase SHIP2 and lamellipodin to mediate the local production of phosphatidylinositol-3,4-bisphosphate and endophilin pre-enrichment. Membrane-bound GTP-loaded Cdc42 recruits FBP17 and CIP4, before being locally deactivated by RICH1 and SH3BP1 GTPase-activating proteins. This generates the transient assembly and disassembly of endophilin spots, which lasts 5-10 seconds. This mechanism periodically primes patches of the membrane for prompt responses upon FEME activation.
Publication types
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Research Support, Non-U.S. Gov't
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Video-Audio Media
MeSH terms
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Animals
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Carrier Proteins / genetics
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Carrier Proteins / metabolism*
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Cell Membrane / metabolism*
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Endocytosis*
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Fatty Acid-Binding Proteins
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GTPase-Activating Proteins / genetics
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GTPase-Activating Proteins / metabolism
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HEK293 Cells
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Humans
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Intracellular Signaling Peptides and Proteins / genetics
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Intracellular Signaling Peptides and Proteins / metabolism*
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Microtubule-Associated Proteins / genetics
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Microtubule-Associated Proteins / metabolism*
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Minor Histocompatibility Antigens / genetics
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Minor Histocompatibility Antigens / metabolism*
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Phosphatidylinositol Phosphates / metabolism
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Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases / genetics
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Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases / metabolism*
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Protein Binding
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Protein Interaction Domains and Motifs
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Rats
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Signal Transduction
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Time Factors
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cdc42 GTP-Binding Protein / genetics
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cdc42 GTP-Binding Protein / metabolism
Substances
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ARHGAP17 protein, human
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Carrier Proteins
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FNBP1 protein, human
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Fatty Acid-Binding Proteins
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GTPase-Activating Proteins
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Intracellular Signaling Peptides and Proteins
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Membrane Proteins
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Microtubule-Associated Proteins
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Minor Histocompatibility Antigens
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Phosphatidylinositol Phosphates
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RAPH1 protein, human
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SH3BP1 protein, human
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SH3GL1 protein, human
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TRIP10 protein, human
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phosphatidylinositol 3,4-diphosphate
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INPPL1 protein, human
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Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases
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CDC42 protein, human
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cdc42 GTP-Binding Protein