The Tyro3 receptor kinase Axl enhances macropinocytosis of Zaire ebolavirus

J Virol. 2011 Jan;85(1):334-47. doi: 10.1128/JVI.01278-09. Epub 2010 Nov 3.

Abstract

Axl, a plasma membrane-associated Tyro3/Axl/Mer (TAM) family member, is necessary for optimal Zaire ebolavirus (ZEBOV) glycoprotein (GP)-dependent entry into some permissive cells but not others. To date, the role of Axl in virion entry is unknown. The focus of this study was to characterize entry pathways that are used for ZEBOV uptake in cells that require Axl for optimal transduction and to define the role of Axl in this process. Through the use of biochemical inhibitors, interfering RNA (RNAi), and dominant negative constructs, we demonstrate that ZEBOV-GP-dependent entry into these cells occurs through multiple uptake pathways, including both clathrin-dependent and caveola/lipid raft-mediated endocytosis. Other dynamin-dependent and -independent pathways such as macropinocytosis that mediate high-molecular-weight dextran uptake also stimulated ZEBOV-GP entry into these cells, and inhibitors that are known to block macropinocytosis inhibited both dextran uptake and ZEBOV infection. These findings provided strong evidence for the importance of this pathway in filovirus entry. Reduction of Axl expression by RNAi treatment resulted in decreased ZEBOV entry via macropinocytosis but had no effect on the clathrin-dependent or caveola/lipid raft-mediated endocytic mechanisms. Our findings demonstrate for the first time that Axl enhances macropinocytosis, thereby increasing productive ZEBOV entry.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Axl Receptor Tyrosine Kinase
  • Cell Line
  • Chlorocebus aethiops
  • Clathrin / metabolism
  • Ebolavirus / genetics
  • Ebolavirus / metabolism
  • Ebolavirus / pathogenicity*
  • Endocytosis
  • Humans
  • Kidney / cytology
  • Kidney / virology
  • Pinocytosis / physiology*
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism*
  • Receptor Protein-Tyrosine Kinases / genetics
  • Receptor Protein-Tyrosine Kinases / metabolism*
  • Vero Cells
  • Viral Envelope Proteins / metabolism
  • Virus Internalization

Substances

  • Clathrin
  • Proto-Oncogene Proteins
  • Viral Envelope Proteins
  • envelope glycoprotein, Ebola virus
  • Receptor Protein-Tyrosine Kinases
  • TYRO3 protein, human
  • Axl Receptor Tyrosine Kinase