Clathrin mediates integrin endocytosis for focal adhesion disassembly in migrating cells.
Focal adhesion disassembly is regulated by microtubules (MTs) through an unknown mechanism that involves dynamin. To test whether endocytosis may be involved, we interfered with the function of clathrin or its adaptors autosomal recessive hypercholesteremia (ARH) and Dab2 (Disabled-2) and found that both treatments prevented MT-induced focal adhesion disassembly. Surface labeling experiments showed that integrin was endocytosed in an extracellular matrix-, clathrin-, and ARH- and Dab2-dependent manner before entering Rab5 endosomes. Clathrin colocalized with a subset of focal adhesions in an ARH- and Dab2-dependent fashion. Direct imaging showed that clathrin rapidly accumulated on focal adhesions during MT-stimulated disassembly and departed from focal adhesions with integrin upon their disassembly. In migrating cells, depletion of clathrin or Dab2 and ARH inhibited focal adhesion disassembly and decreased the rate of migration. These results show that focal adhesion disassembly occurs through a targeted mechanism involving MTs, clathrin, and specific clathrin adaptors and that direct endocytosis of integrins from focal adhesions mediates their disassembly in migrating cells.
Ezratty
Ellen J
EJ
Department of Pathology and Cell Biology, Columbia University, New York, NY 10032, USA.
Bertaux
Claire
C
Marcantonio
Eugene E
EE
Gundersen
Gregg G
GG
D048868:Adaptor Proteins, Signal Transducing / Q000037:antagonists & inhibitors; D033942:Adaptor Proteins, Vesicular Transport / Q000037:antagonists & inhibitors; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D002465:Cell Movement; D016764:Cell Polarity; D002966:Clathrin / Q000502:physiology*; D004705:Endocytosis; D022001:Focal Adhesions / Q000378:metabolism*; D016023:Integrins / Q000378:metabolism*; D008565:Membrane Proteins / Q000037:antagonists & inhibitors; D051379:Mice; D008870:Microtubules / Q000502:physiology; D041681:NIH 3T3 Cells; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors; D034622:RNA Interference
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D051017:Apoptosis Regulatory Proteins; D002966:Clathrin; C478773:Dab2 protein, mouse; D016023:Integrins; C500241:Ldlrap1 protein, mouse; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C106293:Numb protein, mouse