Entry of Ebolavirus (EBOV) into host cells is mediated by its sole glycoprotein, known as GP. The GP andits associated EBOV entry events possess many unusual features that provide novel insights into ourfundamental understanding of viral entry. In this R21 project, we aim to elucidate how the newly identifiedcellular restriction factors, known as interferon-inducible transmembrane (IFITM) proteins, especially IFITM2,potently and specifically inhibit EBOV entry, and in doing so, aid the development of novel antiviraltherapeutics.
Aim 1 : Establish a single virus fusion assay for EBOV and dissect the stages of membrane fusioninhibited by IFITM2. We will take advantage of the fact that EBOV GP can be efficiently incorporated into itsvirus-like particles (VLPs) formed by the VP40 matrix protein, and develop a single virus imaging and fusionsystem to determine how IFITM2 inhibits EBOV fusion in endolysosomes.
Aim 2 : Elucidate the molecular andbiochemical mechanisms by which IFITM2 specifically inhibits EBOV GP-mediated entry. We will test the centralhypothesis that IFITM2 profoundly inhibits EBOV entry by disturbing the triggering capability and/or thecholesterol transport activity of its intracellular receptor, Niemann-Pick C1 (NPC1). A series of biochemical andnovel fluorescence lipid labeling techniques will be used to assess the effect of IFITM2 on cholesterol content,membrane fluidity, and conformational changes of EBOV GP. EBOV is a highly pathogenic filovirus thatcauses severe hemorrhagic fever in humans, with a fatality rate of up to 90%. Results from the proposedstudies will provide critical novel insight into how IFITM2 restricts EBOV GP-mediated membrane fusion andentry, as well as advance our understanding of the general mechanism of IFITMs that block viral entry.
Ebola virus causes fatal hemorrhagic fever in human and non-human primates. Currently; no effective drug orFDA-approved vaccine is available for this deadly virus. We study how some cellular factors intrinsically inhibitEbola virus entry into host cells; which may lead to new strategies to prevent and treat infection by Ebola.
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