Ndm, a coiled-coil domain protein that suppresses macropinocytosis and has effects on cell migration

Mol Biol Cell. 2012 Sep;23(17):3407-19. doi: 10.1091/mbc.E12-05-0392. Epub 2012 Jul 18.

Abstract

The ampA gene has a role in cell migration in Dictyostelium discoideum. Cells overexpressing AmpA show an increase in cell migration, forming large plaques on bacterial lawns. A second-site suppressor of this ampA-overexpressing phenotype identified a previously uncharacterized gene, ndm, which is described here. The Ndm protein is predicted to contain a coiled-coil BAR-like domain-a domain involved in endocytosis and membrane bending. ndm-knockout and Ndm-monomeric red fluorescent protein-expressing cell lines were used to establish a role for ndm in suppressing endocytosis. An increase in the rate of endocytosis and in the number of endosomes was detected in ndm(-) cells. During migration ndm(-) cells formed numerous endocytic cups instead of the broad lamellipodia structure characteristic of moving cells. A second lamellipodia-based function-cell spreading-was also defective in the ndm(-) cells. The increase in endocytosis and the defect in lamellipodia formation were associated with reduced chemotaxis in ndm(-) cells. Immunofluorescence results and glutathione S-transferase pull-down assays revealed an association of Ndm with coronin and F-actin. The results establish ndm as a gene important in regulating the balance between formation of endocytic cups and lamellipodia structures.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 4-Butyrolactone / analogs & derivatives
  • 4-Butyrolactone / metabolism
  • Actin Cytoskeleton / ultrastructure
  • Actins / metabolism
  • Cell Line
  • Cell Movement*
  • Dictyostelium / genetics
  • Dictyostelium / metabolism
  • Dictyostelium / physiology*
  • Dictyostelium / ultrastructure
  • Gene Knockout Techniques
  • Metalloendopeptidases / genetics
  • Metalloendopeptidases / metabolism
  • Pinocytosis*
  • Protein Structure, Tertiary
  • Protozoan Proteins / chemistry*
  • Protozoan Proteins / genetics
  • Protozoan Proteins / metabolism*
  • Pseudopodia / metabolism
  • Pseudopodia / ultrastructure

Substances

  • Actins
  • Protozoan Proteins
  • coronin
  • AmpA protein, Dictyostelium protozoan
  • Metalloendopeptidases
  • 4-Butyrolactone