Structure-function analysis of the determinants of CD46 polarization in T cells-combined role for polarity proteins,lipid rafts and ERM proteins (87.19)

CD46 is a ubiquitously expressed human cell surface protein that acts as a receptor for complements for various pathogens including the measles virus. We demonstrated that ligation of the immunoregulatory cell surface receptor, CD46, altered T cell polarity and impaired activation and effector function in response to TCR or NK cell receptor signalling. However the molecular mechanisms by which T cell function is inhibited are not known. We have previously shown that CD46 binds to the polarity protein, Discs large (Dlg), and that this interaction is important for the polarized localization of CD46. Specifically, CD46 localizes to the uropod of T cells, and to the distal pole of T cells undergoing antigen presentation. Polarization of CD46 is partially reduced by mutation of the Dlg-binding site, and is also partially reduced by mutation of the Cysteine residue in the transmembrane domain that is palmitoylated to allow association of CD46 with lipid rafts. However, comparison of CD46 mutants suggests that other determinants are also important in CD46 polarization. We hypothesize that the ERM proteins interacts with CD46, regulates its polarization in T cells, and perhaps play a role in CD46 signal transduction. To assess this we have generated mutations in the ERM-binding consensus sequences of CD46 and expressed these mutants in a uropod-containing T cell line for functional characterization. These studies will help to elucidate the molecular mechanisms by which CD46 exerts its immunoregulatory effects.