An evolutionary-conserved VPS34-PIKfyve-TRPML1-Myosin II axis regulates the speed of amoeboid cell migration

Abstract

Amoeboid cell migration is key to efficient T cell immunity. Spatial polarization of organelles within cells, including endo-lysosomes, is a prerequisite of migration. However, how ultrastructural polarization is linked to the signaling requirements governing T cell migration, remains unknown. Here we show that signaling molecules generated by endo-lysosome-localized kinases regulate velocity of amoeboid migration. Specifically, imaging of T cells identified accumulation of endo-lysosomes decorated with the lipid kinases VPS34-PIKfyve at the uropod of polarized cells. Activity of VPS34 and PIKfyve regulated speed, but not directedness, of migrating T cells. Mechanistically, PI(3,5)P2 generated by the sequential action of VPS34 and PIKfyve mediated Ca2+ efflux from lysosomes via the mucolipin TRP cation channel 1 (TRPML1), thus controlling activity of myosin IIA and hence the generation of propulsive force through retrograde actin flow. The VPS34-PIKfyve kinases also regulated velocity of myeloid cells, as well as of the amoeba Dictyostelium discoideum - establishing the axis as an evolutionary conserved speed control system of amoeboid cell migration. Graphical AbstractO_LIThe VPS34-PIKfyve axis is active on endo-lysosomes at the uropod of migrating T cells. C_LIO_LIVPS34 and PIKfyve promote myosin IIA activation and retrograde action flow. C_LIO_LIAmoeboid cell migration speed is controlled by VPS34 and PIKfyve via TRPML1. C_LIO_LIRegulation of amoeboid migration speed is a conserved function of the VPS34-PIKfyve axis. C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=83 SRC="FIGDIR/small/575998v1_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@1b70e64org.highwire.dtl.DTLVardef@111d052org.highwire.dtl.DTLVardef@83bc24org.highwire.dtl.DTLVardef@b51931_HPS_FORMAT_FIGEXP M_FIG C_FIG