The role of endocytosis in regulating L1-mediated adhesion.
L1 is a neural cell adhesion molecule critical for neural development. Full-length L1 (L1(FL)) contains an alternatively spliced cytoplasmic sequence, RSLE, which is absent in L1 expressed in nonneuronal cells. The RSLE sequence follows a tyrosine, creating an endocytic motif that allows rapid internalization via clathrin-mediated endocytosis. We hypothesized that L1(FL) would internalize more rapidly than L1 lacking the RSLE sequence (L1(Delta)(RSLE)) and that internalization might regulate L1-mediated adhesion. L1 internalization was measured by immunofluorescence microscopy and by uptake of (125)I-anti-rat-L1 antibody, demonstrating that L1(FL) is internalized 2-3 times faster than L1(Delta)(RSLE). Inhibition of clathrin-mediated endocytosis slowed internalization of L1(FL) but did not affect initial uptake of L1(Delta)(RSLE). To test whether L1 endocytosis regulates L1 adhesion, cell aggregation rates were tested. L1(Delta)(RSLE) cells aggregated two times faster than L1(FL) cells. Inhibition of clathrin-mediated endocytosis increases the aggregation rate of the L1(FL) cells to that of L1(Delta)(RSLE) cells. Our results demonstrate that rapid internalization of L1 dramatically affects L1 adhesion.
Long
K E
KE
Department of Neurosciences, Case Western Reserve University, Cleveland, Ohio 44106, USA.
Asou
H
H
Snider
M D
MD
Lemmon
V
V
D000818:Animals; D000954:Antigens, Surface / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology*; D004705:Endocytosis / Q000502:physiology*; D007150:Immunohistochemistry; D007700:Kinetics; D007739:L Cells; D039841:Leukocyte L1 Antigen Complex; D008562:Membrane Glycoproteins / Q000502:physiology*; D051379:Mice; D019006:Neural Cell Adhesion Molecules / Q000502:physiology*; D011188:Potassium / Q000494:pharmacology
D000954:Antigens, Surface; D039841:Leukocyte L1 Antigen Complex; D008562:Membrane Glycoproteins; D019006:Neural Cell Adhesion Molecules; D011188:Potassium