Membrane phospholipid dynamics during cytokinesis: regulation of actin filament assembly by redistribution of membrane surface phospholipid

Abstract

To study molecular motion and function of membrane phospholipids, we have developed various probes which bind specifically to certain phospholipids. Using a novel peptide probe, Ro09-0198, which binds specifically to phosphatidylethanolamine (PE) in biological membranes, we have analyzed the cell surface movement of PE in dividing CHO cells. We found that PE was exposed on the cell surface specifically at the cleavage furrow during the late telophase of cytokinesis. PE was exposed on the cell surface only during the late telophase and no alteration in the distribution of the plasma membranebound peptide was observed during the cytokinesis, suggesting that the surface exposure of PE reflects the enhanced transbilayer movement of PE at the cleavage furrow. Furthermore, cell surface immobilization of PE induced by adding of the cyclic peptide coupled with streptavidin to prometaphase cells effectively blocked the cytokinesis at late telophase. The peptide–streptavidin complex bound specifically to cleavage furrow and inhibited both actin filament disassembly at cleavage furrow and subsequent plasma membrane fusion. Binding of the peptide complex to interphase cells also induced immediate disassembly of stress fibers followed by assembly of cortical actin filaments to the local area of plasma membrane where the peptide complex bound. The cytoskeletal reorganizations caused by the peptide complex were fully reversible; removal of the surface-bound peptide complex by incubating with PE-containing liposome caused gradual disassembly of the cortical actin filaments and subsequent formation of stress fibers. These observations suggest that the redistribution of plasma membrane phospholipids act as a regulator of actin cytoskeleton organization and may play a crucial role in mediating a coordinate movement between plasma membrane and actin cytoskeleton to achieve successful cell division.