Palmitoylation supports assembly and function of integrin-tetraspanin complexes.
As observed previously, tetraspanin palmitoylation promotes tetraspanin microdomain assembly. Here, we show that palmitoylated integrins (alpha3, alpha6, and beta4 subunits) and tetraspanins (CD9, CD81, and CD63) coexist in substantially overlapping complexes. Removal of beta4 palmitoylation sites markedly impaired cell spreading and signaling through p130Cas on laminin substrate. Also in palmitoylation-deficient beta4, secondary associations with tetraspanins (CD9, CD81, and CD63) were diminished and cell surface CD9 clustering was decreased, whereas core alpha6beta4-CD151 complex formation was unaltered. There is also a functional connection between CD9 and beta4 integrins, as evidenced by anti-CD9 antibody effects on beta4-dependent cell spreading. Notably, beta4 palmitoylation neither increased localization into "light membrane" fractions of sucrose gradients nor decreased solubility in nonionic detergents-hence it does not promote lipid raft association. Instead, palmitoylation of beta4 (and of the closely associated tetraspanin CD151) promotes CD151-alpha6beta4 incorporation into a network of secondary tetraspanin interactions (with CD9, CD81, CD63, etc.), which provides a novel framework for functional regulation.
Yang
Xiuwei
X
Dana-Farber Cancer Institute and Department of Pathology, Harvard Medical School, Boston, MA 02115, USA.
Kovalenko
Oleg V
OV
Tang
Wei
W
Claas
Christoph
C
Stipp
Christopher S
CS
Hemler
Martin E
ME
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002460:Cell Line; D006801:Humans; D039661:Integrin beta3 / Q000378:metabolism; D016023:Integrins / Q000378:metabolism*; D046911:Macromolecular Substances / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D021962:Membrane Microdomains / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D019308:Palmitic Acid / Q000378:metabolism*; D021122:Protein Subunits / Q000378:metabolism; D017978:Receptors, Growth Factor / Q000378:metabolism; D060245:Tetraspanin 29; D019061:src-Family Kinases / Q000378:metabolism
D015703:Antigens, CD; C554869:CD9 protein, human; D039661:Integrin beta3; D016023:Integrins; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D021122:Protein Subunits; D017978:Receptors, Growth Factor; D060245:Tetraspanin 29; D019308:Palmitic Acid; D019061:src-Family Kinases