Phospholipase D2 is required for efficient endocytic recycling of transferrin receptors.

RNA interference-mediated depletion of phospholipase D2 (PLD2), but not PLD1, inhibited recycling of transferrin receptors in HeLa cells, whereas the internalization rate was unaffected by depletion of either PLD. Although reduction of both PLD isoforms inhibits PLD activity stimulated by phorbol 12-myristic 13-acetate, only depletion of PLD2 decreased nonstimulated activity. Cells with reduced PLD2 accumulated a greater fraction of transferrin receptors in a perinuclear compartment that was positive for Rab11, a marker of recycling endosomes.

Differently anchored influenza hemagglutinin mutants display distinct interaction dynamics with mutual rafts.

Lipid rafts play important roles in cellular functions through concentrating or sequestering membrane proteins. This requires proteins to differ in the stability of their interactions with lipid rafts. However, knowledge of the dynamics of membrane protein-raft interactions is lacking.

Features of influenza HA required for apical sorting differ from those required for association with DRMs or MAL.

The influenza virus hemagglutinin (HA) is sorted to the apical membrane in polarized epithelial cells and associates with detergent-resistant membranes (DRMs). By systematic mutagenesis of the transmembrane residues, we show that hemagglutinin requires 10 contiguous transmembrane amino acids to enter detergent-resistant membranes and that the surface of the trimeric hemagglutinin transmembrane domain facing the lipid environment as well as that facing the interior of the trimer is important for stable association with detergent-resistant membranes. However, association with detergent-resistant membranes was not required for apical sorting.