Integrin-beta3 clusters recruit clathrin-mediated endocytic machinery in the absence of traction force.
The turnover of integrin receptors is critical for cell migration and adhesion dynamics. Here we find that force development at integrins regulates adaptor protein recruitment and endocytosis. Using mobile RGD (Arg-Gly-Asp) ligands on supported lipid membranes (RGD membranes) and rigid RGD ligands on glass (RGD-glass), we find that matrix force-dependent integrin signals block endocytosis. Dab2, an adaptor protein of clathrin-mediated endocytosis, is not recruited to activated integrin-beta3 clusters on RGD-glass; however, it is recruited to integrin-mediated adhesions on RGD membranes. Further, when force generation is inhibited on RGD-glass, Dab2 binds to integrin-beta3 clusters. Dab2 binding to integrin-beta3 excludes other adhesion-related adaptor proteins, such as talin. The clathrin-mediated endocytic machinery combines with Dab2 to facilitate the endocytosis of RGD-integrin-beta3 clusters. From these observations, we propose that loss of traction force on ligand-bound integrin-beta3 causes recruitment of Dab2/clathrin, resulting in endocytosis of integrins.
School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.|Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.
Cheng-han
http://orcid.org/0000-0002-8821-8877
CH
Yu
Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.|Randall Division of Cell and Molecular Biophysics, King's College London, London SE1 1UL, UK.
Nisha Bte Mohd
NB
Rafiq
School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
Fakun
F
Cao
School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
Yuhuan
Y
Zhou
Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.
Anitha
A
Krishnasamy
Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.
Kabir Hassan
KH
Biswas
Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.
Andrea
A
Ravasio
Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.
Zhongwen
Z
Chen
Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.
Yu-Hsiu
YH
Wang
Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.|Frontiers of Innovative Research in Science and Technology, Konan University, Kobe 650-0047, Japan.
Keiko
K
Kawauchi
Randall Division of Cell and Molecular Biophysics, King's College London, London SE1 1UL, UK.
Gareth E
http://orcid.org/0000-0001-5879-3048
GE
Jones
Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.|Department of Biological Sciences, Columbia University, New York, New York 10027, USA.
Michael P
MP
Sheetz
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics / Q000378:metabolism; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D001696:Biomechanical Phenomena; D002465:Cell Movement; D002477:Cells / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism*; D002966:Clathrin / Q000235:genetics / Q000378:metabolism*; D004705:Endocytosis; D006367:HeLa Cells; D006801:Humans; D039661:Integrin beta3 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding; D014143:Traction
D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D051017:Apoptosis Regulatory Proteins; D002966:Clathrin; C478773:Dab2 protein, mouse; D039661:Integrin beta3