Cytoskeleton interactions involved in the assembly and function of glycoprotein-80 adhesion complexes in dictyostelium.
Adhesion complexes typically assemble from clustered receptors that link to the cytoskeleton via cytoplasmic adapter proteins. However, it is unclear how phospholipid-anchored adhesion molecules, such as the Dictyostelium receptor gp80, interact with the cytoskeleton. gp80 has been found to form adhesion complexes from raftlike membrane domains, which can be isolated as a Triton X-100-insoluble floating fraction (TIFF). We report here that the actin-binding protein ponticulin mediates TIFF-cytoskeleton interactions. Analysis of gp80-null cells revealed that these interactions were minimal in the absence of gp80. During development, gp80 was required to enhance these interactions as its adhesion complexes assembled. Whereas ponticulin and gp80 could partition independently into TIFF, gp80 was shown to recruit ponticulin to cell-cell contacts and to increase its partitioning into TIFF. However, these proteins did not co-immunoprecipitate. Furthermore, sterol sequestration abrogated the association of ponticulin with TIFF without affecting gp80, suggesting that sterols may mediate the interactions between ponticulin and gp80. In ponticulin-null cells, large gp80 adhesion complexes assembled in the absence of ponticulin despite the lack of cytoskeleton association. We propose that such nascent gp80 adhesion complexes produce expanded raftlike domains that recruit ponticulin and thereby establish stable cytoskeleton interactions to complete the assembly process.
Harris
Tony J C
TJ
Banting and Best Department of Medical Research, University of Toronto, Ontario M5G 1L6, Canada.
Ravandi
Amir
A
Awrey
Donald E
DE
Siu
Chi-Hung
CH
D000199:Actins / Q000378:metabolism; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002352:Carrier Proteins / Q000494:pharmacology; D002448:Cell Adhesion; D002450:Cell Communication; D003599:Cytoskeleton / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology; D004023:Dictyostelium / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D021902:Enterobacter aerogenes / Q000378:metabolism; D050356:Lipid Metabolism; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000494:pharmacology; D008840:Microfilament Proteins / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D017830:Octoxynol / Q000494:pharmacology; D011233:Precipitin Tests; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines
D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D002352:Carrier Proteins; D003902:Detergents; C034037:F-actin-binding proteins; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D013844:Thiazoles; D053778:Thiazolidines; D017830:Octoxynol; C037068:latrunculin B