Journal article
Characterization of Human and Murine T-Cell Immunoglobulin Mucin Domain 4 (TIM-4) IgV Domain Residues Critical for Ebola Virus Entry
Journal of virology, Vol.90(13), pp.6097-6111
07/01/2016
DOI: 10.1128/JVI.00100-16
PMCID: PMC4907230
PMID: 27122575
Abstract
Phosphatidylserine (PtdSer) receptors that are responsible for the clearance of dying cells have recently been found to mediate enveloped virus entry. Ebola virus (EBOV), a member of the Filoviridae family of viruses, utilizes PtdSer receptors for entry into target cells. The PtdSer receptors human and murine T-cell immunoglobulin mucin (TIM) domain proteins TIM-1 and TIM-4 mediate filovirus entry by binding to PtdSer on the virion surface via a conserved PtdSer binding pocket within the amino-terminal IgV domain. While the residues within the TIM-1 IgV domain that are important for EBOV entry are characterized, the molecular details of virion-TIM-4 interactions have yet to be investigated. As sequences and structural alignments of the TIM proteins suggest distinct differences in the TIM-1 and TIM-4 IgV domain structures, we sought to characterize TIM-4 IgV domain residues required for EBOV entry. Using vesicular stomatitis virus pseudovirions bearing EBOV glycoprotein (EBOV GP/VSVΔG), we evaluated virus binding and entry into cells expressing TIM-4 molecules mutated within the IgV domain, allowing us to identify residues important for entry. Similar to TIM-1, residues in the PtdSer binding pocket of murine and human TIM-4 (mTIM-4 and hTIM-4) were found to be important for EBOV entry. However, additional TIM-4-specific residues were also found to impact EBOV entry, with a total of 8 mTIM-4 and 14 hTIM-4 IgV domain residues being critical for virion binding and internalization. Together, these findings provide a greater understanding of the interaction of TIM-4 with EBOV virions.
With more than 28,000 cases and over 11,000 deaths during the largest and most recent Ebola virus (EBOV) outbreak, there has been increased emphasis on the development of therapeutics against filoviruses. Many therapies under investigation target EBOV cell entry. T-cell immunoglobulin mucin (TIM) domain proteins are cell surface factors important for the entry of many enveloped viruses, including EBOV. TIM family member TIM-4 is expressed on macrophages and dendritic cells, which are early cellular targets during EBOV infection. Here, we performed a mutagenesis screening of the IgV domain of murine and human TIM-4 to identify residues that are critical for EBOV entry. Surprisingly, we identified more human than murine TIM-4 IgV domain residues that are required for EBOV entry. Defining the TIM IgV residues needed for EBOV entry clarifies the virus-receptor interactions and paves the way for the development of novel therapeutics targeting virus binding to this cell surface receptor.
Details
- Title: Subtitle
- Characterization of Human and Murine T-Cell Immunoglobulin Mucin Domain 4 (TIM-4) IgV Domain Residues Critical for Ebola Virus Entry
- Creators
- Bethany A Rhein - Department of Microbiology, The University of Iowa, Iowa City, Iowa, USARachel B Brouillette - Department of Microbiology, The University of Iowa, Iowa City, Iowa, USAGrace A Schaack - Department of Microbiology, The University of Iowa, Iowa City, Iowa, USAJohn A Chiorini - Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USAWendy Maury - Department of Microbiology, The University of Iowa, Iowa City, Iowa, USA wendy-maury@uiowa.edu
- Resource Type
- Journal article
- Publication Details
- Journal of virology, Vol.90(13), pp.6097-6111
- DOI
- 10.1128/JVI.00100-16
- PMID
- 27122575
- PMCID
- PMC4907230
- ISSN
- 0022-538X
- eISSN
- 1098-5514
- Grant note
- U54 AI057160 / NIAID NIH HHS R01 AI077519 / NIAID NIH HHS
- Language
- English
- Date published
- 07/01/2016
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984083889502771
Metrics
27 Record Views