Interaction of the Wiskott–Aldrich syndrome protein with sorting nexin 9 is required for CD28 endocytosis and cosignaling in T cells

K Badour, MKH McGavin, J Zhang… - Proceedings of the …, 2007 - National Acad Sciences
K Badour, MKH McGavin, J Zhang, S Freeman, C Vieira, D Filipp, M Julius, GB Mills
Proceedings of the National Academy of Sciences, 2007National Acad Sciences
The Wiskott–Aldrich syndrome protein (WASp) plays a major role in coupling T cell antigen
receptor (TCR) stimulation to induction of actin cytoskeletal changes required for T cell
activation. Here, we report that WASp inducibly binds the sorting nexin 9 (SNX9) in T cells
and that WASp, SNX9, p85, and CD28 colocalize within clathrin-containing endocytic
vesicles after TCR/CD28 costimulation. SNX9, implicated in clathrin-mediated endocytosis,
binds WASp via its SH3 domain and uses its PX domain to interact with the phosphoinositol …
The Wiskott–Aldrich syndrome protein (WASp) plays a major role in coupling T cell antigen receptor (TCR) stimulation to induction of actin cytoskeletal changes required for T cell activation. Here, we report that WASp inducibly binds the sorting nexin 9 (SNX9) in T cells and that WASp, SNX9, p85, and CD28 colocalize within clathrin-containing endocytic vesicles after TCR/CD28 costimulation. SNX9, implicated in clathrin-mediated endocytosis, binds WASp via its SH3 domain and uses its PX domain to interact with the phosphoinositol 3-kinase regulatory subunit p85 and product, phosphoinositol (3,4,5)P3. The data reveal ligation-induced CD28 endocytosis to be clathrin- and phosphoinositol 3-kinase-dependent and TCR/CD28-evoked CD28 internalization and NFAT activation to be markedly enhanced by SNX9 overexpression, but severely impaired by expression of an SNX9 mutant (SNX9ΔPX) lacking p85-binding capacity. CD28 endocytosis and CD28-evoked actin polymerization also are impaired in WASp-deficient T cells. These findings suggest that SNX9 couples WASp to p85 and CD28 so as to link CD28 engagement to its internalization and to WASp-mediated actin remodeling required for CD28 cosignaling. Thus, the WASp/SNX9/p85/CD28 complex enables a unique interface of endocytic, actin polymerizing, and signal transduction pathways required for CD28-mediated T cell costimulation.
National Acad Sciences