Antagonistic and agonistic effects of an extracellular fragment of nectin on formation of E-cadherin-based cell-cell adhesion.
BACKGROUND
Nectin is a Ca2+-independent immunoglobulin-like cell-cell adhesion molecule at the E-cadherin-based cell-cell adherens junctions (AJs), and comprises a family consisting of four members, nectin-1, -2, -3, and -4. Nectin and E-cadherin are associated with afadin and alpha-catenin, actin filament (F-actin)-binding proteins connecting respective adhesion molecules to the actin cytoskeleton, but the role of nectin in the formation of the E-cadherin-based cell-cell AJs has not yet been fully understood. To obtain evidence for this role of nectin, we attempted to develop an antagonist and/or agonist of nectin.


RESULTS
We made a recombinant extracellular fragment of nectin-3 (Nef-3). Nef-3 trans-interacted with cellular nectin-1 and thereby diminished the formation of the nectin-1-based cell-cell adhesion. This resulted in a reduction of the formation of the E-cadherin-based cell-cell adhesion in L fibroblasts stably expressing both exogenous nectin-1alpha and E-cadherin (nectin-1-EL cells) and MDCK cells stably expressing exogenous nectin-1alpha (nectin-1-MDCK cells). This antagonistic effect of Nef-3 was also observed in L cells stably expressing exogenous E-cadherin alone (EL cells) and wild-type MDCK cells. Conversely, Nef-3 coated on microbeads first recruited the nectin-afadin complex and then the E-cadherin-catenin complex to the bead-cell contact sites in nectin-1-EL and nectin-1-MDCK cells.


CONCLUSIONS
These results suggest that nectin is necessary and sufficient for the recruitment of E-cadherin to the nectin-based cell-cell adhesion sites and involved in the formation of E-cadherin-based cell-cell AJs.
Honda
Tomoyuki
T
Department of Molecular Biology and Biochemistry, Osaka University Graduate School of Medicine/Faculty of Medicine, Suita 565-0871, Japan.
Shimizu
Kazuya
K
Kawakatsu
Tomomi
T
Yasumi
Masato
M
Shingai
Tatsushi
T
Fukuhara
Atsunori
A
Ozaki-Kuroda
Kumi
K
Irie
Kenji
K
Nakanishi
Hiroyuki
H
Takai
Yoshimi
Y
D000818:Animals; D015820:Cadherins / Q000502:physiology*; D002448:Cell Adhesion / Q000187:drug effects* / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000494:pharmacology*; D002478:Cells, Cultured; D004285:Dogs; D007365:Intercellular Junctions / Q000187:drug effects* / Q000502:physiology; D008840:Microfilament Proteins / Q000378:metabolism; D008954:Models, Biological; D000075983:Nectins; D010446:Peptide Fragments / Q000494:pharmacology
D015820:Cadherins; D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; D000075983:Nectins; D010446:Peptide Fragments; C108835:afadin