To elucidate the antiviral mechanism of S-palmitoylated IFITMs (1, 2 and 3) in host immunity against viruses, we propose to perform detailed biochemical, biophysical and cellular studies of these lipidated immune effectors in this grant renewal. For biochemical and structural studies of IFITMs, we will perform computational modeling and reconstitute site-specifically lipidated IFITM3 in vitro (Aim 1). Moreover, we have discovered direct interactions of IFITMs with specific cellular lipids and will investigate their significance biochemically in vitro and in mammalian cells during virus infections (Aim 2). Lastly, we will investigate S-palmitoylated IFITM interactions with key cellular factors involved their regulation and antiviral activity (Aim 3). These studies should help determine how IFITMs prevent virus entry and elucidate biochemical mechanisms by which site-specific lipidation controls membrane protein structure and function in host immunity. Determining the mechanisms that control S-palmitoylated IFITMs function is crucial for understanding host immunity and may reveal new strategies for combatting virus infection in humans.
Interferon-induced transmembrane proteins (IFITMs) are lipidated innate immune proteins that prevent diverse viruses (influenza A virus, Ebola virus, Zika virus and others) from infecting many vertebrates, including humans. While the expression and localization of IFITMs are known to be crucial for restricting virus entry into host cells, the underlying molecular mechanisms are still unclear. To address the antiviral mechanism of IFITMs, this proposal will employ innovative computational, chemical, structural and cellular methods to investigate IFITMs in vitro and in mammalian cells.
| Thinon, Emmanuelle; Fernandez, Joseph P; Molina, Henrik et al. (2018) Selective Enrichment and Direct Analysis of Protein S-Palmitoylation Sites. J Proteome Res 17:1907-1922 |
| Percher, Avital; Thinon, Emmanuelle; Hang, Howard (2017) Mass-Tag Labeling Using Acyl-PEG Exchange for the Determination of Endogenous Protein S-Fatty Acylation. Curr Protoc Protein Sci 89:14.17.1-14.17.11 |
| McMichael, Temet M; Zhang, Lizhi; Chemudupati, Mahesh et al. (2017) The palmitoyltransferase ZDHHC20 enhances interferon-induced transmembrane protein 3 (IFITM3) palmitoylation and antiviral activity. J Biol Chem 292:21517-21526 |
| Westcott, Nathan P; Fernandez, Joseph P; Molina, Henrik et al. (2017) Chemical proteomics reveals ADP-ribosylation of small GTPases during oxidative stress. Nat Chem Biol 13:302-308 |
| Thinon, Emmanuelle; Percher, Avital; Hang, Howard C (2016) Bioorthogonal Chemical Reporters for Monitoring Unsaturated Fatty-Acylated Proteins. Chembiochem 17:1800-1803 |
| Peng, Tao; Hang, Howard C (2016) Site-Specific Bioorthogonal Labeling for Fluorescence Imaging of Intracellular Proteins in Living Cells. J Am Chem Soc 138:14423-14433 |
| Percher, Avital; Ramakrishnan, Srinivasan; Thinon, Emmanuelle et al. (2016) Mass-tag labeling reveals site-specific and endogenous levels of protein S-fatty acylation. Proc Natl Acad Sci U S A 113:4302-7 |
| Peng, Tao; Thinon, Emmanuelle; Hang, Howard C (2016) Proteomic analysis of fatty-acylated proteins. Curr Opin Chem Biol 30:77-86 |
| Thinon, Emmanuelle; Hang, Howard C (2015) Chemical reporters for exploring protein acylation. Biochem Soc Trans 43:253-61 |
| Moudgil, Devinderjit K; Westcott, Nathan; Famulski, Jakub K et al. (2015) A novel role of farnesylation in targeting a mitotic checkpoint protein, human Spindly, to kinetochores. J Cell Biol 208:881-96 |
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