E-cadherin junction formation involves an active kinetic nucleation process.
Epithelial (E)-cadherin-mediated cell-cell junctions play important roles in the development and maintenance of tissue structure in multicellular organisms. E-cadherin adhesion is thus a key element of the cellular microenvironment that provides both mechanical and biochemical signaling inputs. Here, we report in vitro reconstitution of junction-like structures between native E-cadherin in living cells and the extracellular domain of E-cadherin (E-cad-ECD) in a supported membrane. Junction formation in this hybrid live cell-supported membrane configuration requires both active processes within the living cell and a supported membrane with low E-cad-ECD mobility. The hybrid junctions recruit α-catenin and exhibit remodeled cortical actin. Observations suggest that the initial stages of junction formation in this hybrid system depend on the trans but not the cis interactions between E-cadherin molecules, and proceed via a nucleation process in which protrusion and retraction of filopodia play a key role.
Mechanobiology Institute, National University of Singapore, Singapore 117411;
Kabir H
KH
Biswas
Mechanobiology Institute, National University of Singapore, Singapore 117411; Department of Chemistry, University of California, Berkeley, CA 94720;
Kevin L
KL
Hartman
Mechanobiology Institute, National University of Singapore, Singapore 117411;
Cheng-han
CH
Yu
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032; Howard Hughes Medical Institute, Columbia University, New York, NY 10032; Center for Computational Biology and Bioinformatics, Columbia University, New York, NY 10032;
Oliver J
OJ
Harrison
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032; Howard Hughes Medical Institute, Columbia University, New York, NY 10032; Center for Computational Biology and Bioinformatics, Columbia University, New York, NY 10032;
Hang
H
Song
Howard Hughes Medical Institute, University of California, Berkeley, CA 94720;
Adam W
AW
Smith
Department of Chemistry, University of California, Berkeley, CA 94720;
William Y C
WY
Huang
Howard Hughes Medical Institute, University of California, Berkeley, CA 94720;
Wan-Chen
WC
Lin
Mechanobiology Institute, National University of Singapore, Singapore 117411;
Zhenhuan
Z
Guo
Mechanobiology Institute, National University of Singapore, Singapore 117411;
Anup
A
Padmanabhan
Department of Dermatology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611;
Sergey M
SM
Troyanovsky
Kennedy Institute, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, Headington OX3 7FY, United Kingdom;
Michael L
ML
Dustin
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032; Center for Computational Biology and Bioinformatics, Columbia University, New York, NY 10032;
Lawrence
L
Shapiro
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY 10032; Howard Hughes Medical Institute, Columbia University, New York, NY 10032; Center for Computational Biology and Bioinformatics, Columbia University, New York, NY 10032; bh6@cumc.columbia.edu biezbr@nus.edu.sg jtgroves@lbl.gov.
Barry
B
Honig
Mechanobiology Institute, National University of Singapore, Singapore 117411; Department of Biomedical Engineering, National University of Singapore, Singapore 117411; bh6@cumc.columbia.edu biezbr@nus.edu.sg jtgroves@lbl.gov.
Ronen
R
Zaidel-Bar
Mechanobiology Institute, National University of Singapore, Singapore 117411; Department of Chemistry, University of California, Berkeley, CA 94720; Howard Hughes Medical Institute, University of California, Berkeley, CA 94720; Physical Biosciences and Materials Sciences Divisions, Lawrence Berkeley National Laboratory, University of California, Berkeley, CA 94720 bh6@cumc.columbia.edu biezbr@nus.edu.sg jtgroves@lbl.gov.
Jay T
JT
Groves
D001703:Biophysics; D015820:Cadherins / Q000378:metabolism*; D002460:Cell Line; D003599:Cytoskeleton / Q000378:metabolism; D006801:Humans; D007365:Intercellular Junctions; D007700:Kinetics; D008051:Lipid Bilayers; D015398:Signal Transduction
D015820:Cadherins; D008051:Lipid Bilayers
adhesion; bilayer; cadherin; diffusion; nucleation