ADP-ribosylation factor-like 4A interacts with Robo1 to promote cell migration by regulating Cdc42 activation

Mol Biol Cell. 2019 Jan 1;30(1):69-81. doi: 10.1091/mbc.E18-01-0001. Epub 2018 Nov 14.

Abstract

Cell migration is a highly regulated event that is initiated by cell membrane protrusion and actin reorganization. Robo1, a single-pass transmembrane receptor, is crucial for neuronal guidance and cell migration. ADP-ribosylation factor (Arf)-like 4A (Arl4A), an Arf small GTPase, functions in cell morphology, cell migration, and actin cytoskeleton remodeling; however, the molecular mechanisms of Arl4A in cell migration are unclear. Here, we report that the binding of Arl4A to Robo1 modulates cell migration by promoting Cdc42 activation. We found that Arl4A interacts with Robo1 in a GTP-dependent manner and that the Robo1 amino acid residues 1394-1398 are required for this interaction. The Arl4A-Robo1 interaction is essential for Arl4A-induced cell migration and Cdc42 activation but not for the plasma membrane localization of Robo1. In addition, we show that the binding of Arl4A to Robo1 decreases the association of Robo1 with the Cdc42 GTPase-activating protein srGAP1. Furthermore, Slit2/Robo1 binding down-regulates the Arl4A-Robo1 interaction in vivo, thus attenuating Cdc42-mediated cell migration. Therefore, our study reveals a novel mechanism by which Arl4A participates in Slit2/Robo1 signaling to modulate cell motility by regulating Cdc42 activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP-Ribosylation Factors / metabolism*
  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Cell Membrane / metabolism
  • Cell Movement*
  • Chlorocebus aethiops
  • Enzyme Activation
  • GTPase-Activating Proteins / metabolism
  • Guanosine Triphosphate / metabolism
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Models, Biological
  • Nerve Tissue Proteins / chemistry
  • Nerve Tissue Proteins / metabolism*
  • Protein Binding
  • Protein Transport
  • Receptors, Immunologic / chemistry
  • Receptors, Immunologic / metabolism*
  • Roundabout Proteins
  • cdc42 GTP-Binding Protein / metabolism*

Substances

  • GTPase-Activating Proteins
  • Intercellular Signaling Peptides and Proteins
  • Nerve Tissue Proteins
  • Receptors, Immunologic
  • SRGAP1 protein, human
  • Guanosine Triphosphate
  • ADP-Ribosylation Factors
  • ARL4A protein, human
  • cdc42 GTP-Binding Protein
  • Slit homolog 2 protein