Amalgam is a ligand for the transmembrane receptor neurotactin and is required for neurotactin-mediated cell adhesion and axon fasciculation in Drosophila.
Neurotactin (NRT), a member of the cholinesterase-homologous protein family, is a heterophilic cell adhesion molecule that is required for proper axon guidance during Drosophila development. In this study, we identify amalgam (AMA), a member of the immunoglobulin superfamily, as a ligand for the NRT receptor. Using transfected Schneider 2 cells and embryonic primary cultures, we demonstrate that AMA is a secreted protein. Furthermore, AMA is necessary for NRT-expressing cells both to aggregate with themselves and to associate with embryonic primary culture cells. Aggregation assays performed with truncated NRT molecules reveal that the integrity of the cholinesterase-like extracellular domain was not required either for AMA binding or for adhesion, with only amino acids 347-482 of the extracellular domain being necessary for both activities. Moreover, the NRT cytoplasmic domain is required for NRT-mediated adhesion, although not for AMA binding. Using an ama-deficient stock, we find that ama function is not essential for viability. Pupae deficient for ama do exhibit defasciculation defects of the ocellar nerves similar to those found in nrt mutants.
Frémion
F
F
Laboratoire de Génétique et de Physiologie du Développement, IBDM, CNRS/INSERM/Université de la Méditerranée/A.P. de Marseille, Parc Scientifique de Luminy Case 907, 13288 Marseille, Cedex 9, France.
Darboux
I
I
Diano
M
M
Hipeau-Jacquotte
R
R
Seeger
M A
MA
Piovant
M
M
D000818:Animals; D001369:Axons; D001483:Base Sequence; D002448:Cell Adhesion; D002478:Cells, Cultured; D017931:DNA Primers; D004330:Drosophila / Q000196:embryology*; D029721:Drosophila Proteins; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D007136:Immunoglobulins / Q000235:genetics / Q000378:metabolism*; D008024:Ligands; D008562:Membrane Glycoproteins / Q000378:metabolism*; D012313:RNA / Q000378:metabolism
D017931:DNA Primers; D029721:Drosophila Proteins; D007136:Immunoglobulins; D008024:Ligands; D008562:Membrane Glycoproteins; C066176:Nrt protein, Drosophila; D012313:RNA