TIM-1 Mediates Dystroglycan-Independent Entry of Lassa Virus

J Virol. 2018 Jul 31;92(16):e00093-18. doi: 10.1128/JVI.00093-18. Print 2018 Aug 15.


Lassa virus (LASV) is an Old World arenavirus responsible for hundreds of thousands of infections in West Africa every year. LASV entry into a variety of cell types is mediated by interactions with glycosyltransferase LARGE-modified O-linked glycans present on the ubiquitous receptor α-dystroglycan (αDG). However, cells lacking αDG are permissive to LASV infection, suggesting that alternative receptors exist. Previous studies demonstrated that the phosphatidylserine (PtdSer)-binding receptors Axl and Tyro3 along with C-type lectin receptors mediate αDG-independent entry. Here, we demonstrate that another PtdSer receptor, TIM-1, mediates LASV glycoprotein (GP)-pseudotyped virion entry into αDG-knocked-out HEK 293T and wild-type (WT) Vero cells, which express αDG lacking appropriate glycosylation. To investigate the mechanism by which TIM-1 mediates enhancement of entry, we demonstrate that mutagenesis of the TIM-1 IgV domain PtdSer-binding pocket abrogated transduction. Furthermore, the human TIM-1 IgV domain-binding monoclonal antibody ARD5 blocked transduction of pseudovirions bearing LASV GP in a dose-dependent manner. Finally, as we showed previously for other viruses that use TIM-1 for entry, a chimeric TIM-1 protein that substitutes the proline-rich region (PRR) from murine leukemia virus envelope (Env) for the mucin-like domain served as a competent receptor. These studies provide evidence that, in the absence of a functional αDG, TIM-1 mediates the entry of LASV pseudoviral particles through interactions of virions with the IgV PtdSer-binding pocket of TIM-1.IMPORTANCE PtdSer receptors, such as TIM-1, are emerging as critical entry factors for many enveloped viruses. Most recently, hepatitis C virus and Zika virus have been added to a growing list. PtdSer receptors engage with enveloped viruses through the binding of PtdSer embedded in the viral envelope, defining them as GP-independent receptors. This GP-independent entry mechanism should effectively mediate the entry of all enveloped viruses, yet LASV GP-pseudotyped viruses were previously found to be unresponsive to PtdSer receptor enhancement in HEK 293T cells. Here, we demonstrate that LASV pseudovirions can utilize the PtdSer receptor TIM-1 but only in the absence of appropriately glycosylated α-dystroglycan (αDG), the high-affinity cell surface receptor for LASV. Our studies shed light on LASV receptor utilization and explain why previous studies performed with α-DG-expressing cells did not find that LASV pseudovirions utilize PtdSer receptors for virus uptake.

Keywords: Lassa virus; TIM-1; arenavirus; phosphatidylserine; phosphatidylserine receptor; receptor; viral lipids; virus entry.

Publication types

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

MeSH terms

  • Animals
  • Chlorocebus aethiops
  • DNA Mutational Analysis
  • Dystroglycans / deficiency*
  • HEK293 Cells
  • Hepatitis A Virus Cellular Receptor 1 / genetics
  • Hepatitis A Virus Cellular Receptor 1 / metabolism*
  • Host-Pathogen Interactions*
  • Humans
  • Lassa virus / physiology*
  • Receptors, Virus / genetics
  • Receptors, Virus / metabolism*
  • Vero Cells
  • Virus Internalization*


  • HAVCR1 protein, human
  • Hepatitis A Virus Cellular Receptor 1
  • Receptors, Virus
  • Dystroglycans