Neuroglian-mediated cell adhesion induces assembly of the membrane skeleton at cell contact sites.
The protein ankyrin links integral membrane proteins to the spectrin-based membrane skeleton. Ankyrin is often concentrated within restricted membrane domains of polarized epithelia and neurons, but the mechanisms responsible for membrane targeting and its segregation within a continuous lipid bilayer remain unexplained. We provide evidence that neuroglian, a cell adhesion molecule related to L1 and neurofascin, can transmit positional information directly to ankyrin and thereby polarize its distribution in Drosophila S2 tissue culture cells. Ankyrin was not normally associated with the plasma membrane of these cells. Upon expression of an inducible neuroglian minigene, however, cells aggregated into large clusters and ankyrin became concentrated at sites of cell-cell contact. Spectrin was also recruited to sites of cell contact in response to neuroglian expression. The accumulation of ankyrin at cell contacts required the presence of the cytoplasmic domain of neuroglian since a glycosyl phosphatidylinositol-linked form of neuroglian failed to recruit ankyrin to sites of cell-cell contact. Double-labeling experiments revealed that, whereas ankyrin was strictly associated with sites of cell-cell contact, neuroglian was more broadly distributed over the cell surface. A direct interaction between neuroglian and ankyrin was demonstrated using yeast two-hybrid analysis. Thus, neuroglian appears to be activated by extracellular adhesion so that ankyrin and the membrane skeleton selectively associate with sites of cell contact and not with other regions of the plasma membrane.
Dubreuil
R R
RR
Department of Pharmacological & Physiological Sciences, University of Chicago, Illinois 60637, USA.
MacVicar
G
G
Dissanayake
S
S
Liu
C
C
Homer
D
D
Hortsch
M
M
D000818:Animals; D017487:Ankyrins / Q000032:analysis; D001483:Base Sequence; D002448:Cell Adhesion / Q000502:physiology; D015816:Cell Adhesion Molecules, Neuronal / Q000032:analysis / Q000502:physiology*; D002478:Cells, Cultured / Q000737:chemistry / Q000166:cytology; D003599:Cytoskeleton / Q000502:physiology*; D004330:Drosophila / Q000166:cytology*; D029721:Drosophila Proteins; D005455:Fluorescent Antibody Technique; D007365:Intercellular Junctions / Q000737:chemistry / Q000502:physiology; D008565:Membrane Proteins / Q000502:physiology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000032:analysis; D013049:Spectrin / Q000032:analysis; D015003:Yeasts / Q000737:chemistry
D017487:Ankyrins; D015816:Cell Adhesion Molecules, Neuronal; D029721:Drosophila Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; D013049:Spectrin; C061745:Nrg protein, Drosophila