Actin sequestering protein, profilin, regulates intracellular vesicle transport in Leishmania

[Display omitted] •Leishmania express single homolog of profilin.•It undergoes phosphorylation during post-translational modification.•It is localized to the cytoplasm, the flagellum, nucleus and kinetoplast in Leishmania cells.•It binds to G-actin, phosphoinositides, PLP stretches, and sequesters G-actin at high concentration.•It regulates intracellular transport in Leishmania promastigotes. Profilins are the key regulators of actin dynamics in all eukaryotic cells. However, little information is available on their biochemical properties and functions in kinetoplastids, such as Trypanosoma and Leishmania. We show here that Leishmania parasites express only one homolog of profilin (LdPfn), which catalyzes nucleotide exchange on G-actin and promotes actin polymerization at its low concentrations. However, at high concentrations, it strongly inhibits the polymerization process by sequestering actin monomers. We further demonstrate that LdPfn binds to actin in Leishmania promastigotes, by both immunofluorescence microscopy and IgG affinity chromatography. Further, we reveal that this protein besides binding to poly-null-proline motifs, also binds more efficiently to PI(3,5)P2, which is found on early or late endosomes or lysosomes, than to PI(4,5)P2 and PI(3,4,5)P3. Additionally, we show that heterozygous mutants of profilin display significantly slower growth and intracellular vesicle trafficking activity, which is reversed on episomal gene complementation. Together, these findings suggest that profilin regulates intracellular vesicle trafficking in Leishmania perhaps through its binding to polyphosphoinositides.