The trafficking protein JFC1 regulates Rac1-GTP localization at the uropod controlling neutrophil chemotaxis and in vivo migration

J Leukoc Biol. 2019 Jun;105(6):1209-1224. doi: 10.1002/JLB.1VMA0818-320R. Epub 2019 Feb 12.

Abstract

Neutrophil chemotaxis is essential in responses to infection and underlies inflammation. In neutrophils, the small GTPase Rac1 has discrete functions at both the leading edge and in the retraction of the trailing structure at the cell's rear (uropod), but how Rac1 is regulated at the uropod is unknown. Here, we identified a mechanism mediated by the trafficking protein synaptotagmin-like 1 (SYTL1 or JFC1) that controls Rac1-GTP recycling from the uropod and promotes directional migration of neutrophils. JFC1-null neutrophils displayed defective polarization and impaired directional migration to N-formyl-methionine-leucyl-phenylalanine in vitro, but chemoattractant-induced actin remodeling, calcium signaling and Erk activation were normal in these cells. Defective chemotaxis was not explained by impaired azurophilic granule exocytosis associated with JFC1 deficiency. Mechanistically, we show that active Rac1 localizes at dynamic vesicles where endogenous JFC1 colocalizes with Rac1-GTP. Super-resolution microscopy (STORM) analysis shows adjacent distribution of JFC1 and Rac1-GTP, which increases upon activation. JFC1 interacts with Rac1-GTP in a Rab27a-independent manner to regulate Rac1-GTP trafficking. JFC1-null cells exhibited Rac1-GTP accumulation at the uropod and increased tail length, and Rac1-GTP uropod accumulation was recapitulated by inhibition of ROCK or by interference with microtubule remodeling. In vivo, neutrophil dynamic studies in mixed bone marrow chimeric mice show that JFC1-/- neutrophils are unable to move directionally toward the source of the chemoattractant, supporting the notion that JFC1 deficiency results in defective neutrophil migration. Our results suggest that defective Rac1-GTP recycling from the uropod affects directionality and highlight JFC1-mediated Rac1 trafficking as a potential target to regulate chemotaxis in inflammation and immunity.

Keywords: JFC1; Rac1; chemotaxis; neutrophils; vesicular trafficking.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chemotaxis / genetics
  • Chemotaxis / immunology*
  • Guanosine Triphosphate / genetics
  • Guanosine Triphosphate / immunology*
  • Inflammation / genetics
  • Inflammation / immunology
  • Inflammation / pathology
  • Membrane Proteins / genetics
  • Membrane Proteins / immunology*
  • Mice
  • Mice, Knockout
  • Neuropeptides / genetics
  • Neuropeptides / immunology*
  • Neutrophils / immunology*
  • Neutrophils / pathology
  • Pseudopodia / genetics
  • Pseudopodia / immunology*
  • Pseudopodia / pathology
  • Vesicular Transport Proteins / genetics
  • Vesicular Transport Proteins / immunology*
  • rab27 GTP-Binding Proteins / genetics
  • rab27 GTP-Binding Proteins / immunology
  • rac1 GTP-Binding Protein / genetics
  • rac1 GTP-Binding Protein / immunology*

Substances

  • Membrane Proteins
  • Neuropeptides
  • Rac1 protein, mouse
  • Sytl1 protein, mouse
  • Vesicular Transport Proteins
  • rab27 GTP-Binding Proteins
  • Guanosine Triphosphate
  • Rab27a protein, mouse
  • rac1 GTP-Binding Protein