Conditional intercellular cohesion in a Dictyostelium discoideum mutant which is temperature sensitive for correct processing of asparagine-linked oligosaccharides.
Mutants of Dictyostelium discoideum have been isolated by a selection for cells with temperature-sensitive defects in the maturation of glycoprotein N-linked oligosaccharides. Here we describe a mutant, HT7, which is unable to aggregate at the restrictive temperature, but which aggregates and makes fruiting bodies at the permissive temperature. HT7 shows normal early developmental intercellular cohesion, but is temperature sensitive for expression of the ethylenediamine-tetraacetic acid (EDTA)-resistant cohesion characteristic of aggregation. The mutant initiates aggregation, but forms only loose cell mounds which later disperse. Metabolic labelling studies indicate that the thermolabile defect is not in protein synthesis, assembly of the lipid-linked precursor of N-linked oligosaccharides or transfer of the precursor to proteins. However, the defect does prevent assembly of fully processed N-linked oligosaccharides. Further, two glycopeptides, obtained from exhaustive Pronase digests of wild-type plasma membrane glycoproteins, inhibit intercellular cohesion of aggregation-stage wild-type cells. HT7 produces only approximately 50% of the wild-type level of these glycopeptides at the restrictive temperature and one of the glycopeptides has reduced cohesion inhibition ability. A revertant of HT7 was found to aggregate normally, to have restored EDTA-resistant cohesion, to have normal profiles of N-linked oligosaccharides and to express the two cohesion-inhibiting glycopeptides normally. These data strongly support a model in which cohesion during late aggregation is at least in part due to recognition between surface glycans and receptors on neighbouring cells.
Boose
J A
JA
Department of Biology, Georgetown University, Washington, DC 20057.
Henderson
E J
EJ
D000371:Agglutination; D000595:Amino Acid Sequence; D000818:Animals; D002240:Carbohydrate Sequence; D000242:Cyclic AMP / Q000378:metabolism; D004023:Dictyostelium / Q000235:genetics* / Q000502:physiology; D005643:Fucose / Q000378:metabolism; D006017:Glycolipids / Q000096:biosynthesis / Q000302:isolation & purification; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D006031:Glycosylation; D008358:Mannose / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D013431:Sulfates / Q000378:metabolism; D013462:Sulfur Radioisotopes; D013696:Temperature; D014316:Tritium
D006017:Glycolipids; D006023:Glycoproteins; D013431:Sulfates; D013462:Sulfur Radioisotopes; D014316:Tritium; D005643:Fucose; D000242:Cyclic AMP; D008358:Mannose