Utrophin-dystroglycan complex in membranes of adherent cultured cells.
In skeletal muscle, dystrophin binds to an oligomeric, transmembrane complex (DAGc; dystrophin-associated glycoprotein complex) which interacts with laminin in the extracellular matrix. We now present biochemical evidence for an association between utrophin (dystrophin-related protein, DRP) and a major DAGc component, beta-dystroglycan (43DAG) in cultured cell lines which contain little if any dystrophin. We have shown also that utrophin and beta-dystroglycan co-localise at or near the plasma membrane and that they co-sediment in large complexes on sucrose density gradients. On the lower plasma membrane, in contact with the substratum, part of the utrophin and beta-dystroglycan staining co-localised with alpha-actinin in a punctate distribution outside classical vinculin-rich focal adhesions. beta-dystroglycan, utrophin, syntrophin (59DAP), and alpha-actinin were found in all adhesion-competent cell lines studied, but levels of the last three proteins were greatly reduced in myeloma cells, which cannot readily attach to substrata. Possible roles for utrophin in cultured cells are considered in the light of recent evidence for involvement of utrophin-glycoprotein complexes in muscle in signal transduction and recruitment of acetylcholine receptors to neuromuscular junctions.
James
M
M
MRIC Biotechnology Group, N.E. Wales Institute, Deeside, Clwyd, UK.
Nguyen
T M
TM
Wise
C J
CJ
Jones
G E
GE
Morris
G E
GE
D000185:Actinin / Q000032:analysis / Q000648:ultrastructure; D000818:Animals; D015153:Blotting, Western; D002448:Cell Adhesion; D002462:Cell Membrane / Q000737:chemistry*; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D003598:Cytoskeletal Proteins / Q000032:analysis* / Q000648:ultrastructure; D049030:Dystroglycans; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000648:ultrastructure; D008565:Membrane Proteins; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D018482:Muscle, Skeletal / Q000737:chemistry; D011817:Rabbits; D051381:Rats; D049411:Utrophin
D003598:Cytoskeletal Proteins; C483081:DAG1 protein, human; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; C483600:Utrn protein, mouse; D049411:Utrophin; D000185:Actinin; D049030:Dystroglycans