TIM-1 Mediates Dystroglycan-Independent Entry of Lassa Virus

• Published in: Journal of virology Vol. 92; no. 16
• Main Authors: Brouillette, Rachel B, Phillips, Elisabeth K, Patel, Radhika, Mahauad-Fernandez, Wadie, Moller-Tank, Sven, Rogers, Kai J, Dillard, Jacob A, Cooney, Ashley L, Martinez-Sobrido, Luis, Okeoma, Chioma, Maury, Wendy
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 15-08-2018
• Subjects: 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; Virus-Cell Interactions; arenavirus; phosphatidylserine; receptor; viral lipids; virus entry; Lassa virus; phosphatidylserine receptor; TIM-1
• ISSN: 0022-538X; 1098-5514
• DOI: 10.1128/jvi.00093-18

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.
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.