Membrane fusion, mediated by viral spike glycoproteins, is a key process in the infection cycle of all enveloped human and animal viruses. The overall goal of this project is to understand the molecular mechanisms of Ebola and human immunodeficiency virus entry by membrane fusion, by taking combined state-of-the-art structural and cell biophysical imaging approaches. Although the crystal structures of the ectodomains of several enveloped virus spike glycoproteins have been determined to high resolution and although we and others have contributed NMR structures of the membrane- interactive parts of some of these proteins over the years, we are only beginning to understand the structural transformations that take place when these protein domains interact with membranes and each other, and how these structures drive membrane fusion. In the previous grant period we have made substantial progress towards elucidating the structures of the fusion loop, membrane proximal, and transmembrane domains of the Ebola virus envelope glycoprotein and their pH-dependencies in membrane environments. We have also discovered that HIV particles bind and fuse preferentially at lipid discontinuities in plasma membranes of infected cells and that this process depends critically on the level of membrane cholesterol. Building on these achievements, we now propose to (1) solve the structure of the Ebola virus fusion loop in interaction with its membrane-proximal and transmembrane domain, (2) determine the intracellular factors that trigger Ebola virus fusion in the late endosome, and (3) determine the role of membrane heterogeneity in forming the fusion pore for HIV entry.

Public Health Relevance

Viral infections cause many devastating diseases in the US and abroad. Many viruses including influenza, Ebola, and human immunodeficiency viruses, are enveloped by a membrane that encloses their hereditary nucleic acids. To propagate these viruses infect cells by fusing their membrane envelopes with a target membrane of the cell either at the cell surface or in an internal organelle, i.e. the endosome. This project aims at discovering the fundamental mechanisms by which Ebola virus and HIV fuse with their respective target membranes. State--of--the--art structural, biophysical, and high- -resolution imaging approaches will be taken to delineate the cell entry mechanisms of these two viruses. Knowledge gained from this research will help to eventually develop new classes of viral entry inhibitors.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI030557-27
Application #
9625810
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Repik, Patricia M
Project Start
1991-09-01
Project End
2022-12-31
Budget Start
2019-01-01
Budget End
2019-12-31
Support Year
27
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Virginia
Department
Physiology
Type
Schools of Medicine
DUNS #
065391526
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Lee, Jinwoo; Nyenhuis, David A; Nelson, Elizabeth A et al. (2017) Structure of the Ebola virus envelope protein MPER/TM domain and its interaction with the fusion loop explains their fusion activity. Proc Natl Acad Sci U S A 114:E7987-E7996
Yang, Sung-Tae; Kreutzberger, Alex J B; Kiessling, Volker et al. (2017) HIV virions sense plasma membrane heterogeneity for cell entry. Sci Adv 3:e1700338
Yang, Sung-Tae; Kreutzberger, Alex J B; Lee, Jinwoo et al. (2016) The role of cholesterol in membrane fusion. Chem Phys Lipids 199:136-143
Liang, Binyong; Tamm, Lukas K (2016) NMR as a tool to investigate the structure, dynamics and function of membrane proteins. Nat Struct Mol Biol 23:468-74
Yang, Sung-Tae; Kiessling, Volker; Tamm, Lukas K (2016) Line tension at lipid phase boundaries as driving force for HIV fusion peptide-mediated fusion. Nat Commun 7:11401
Lee, Jinwoo; Gregory, Sonia M; Nelson, Elizabeth A et al. (2016) The Roles of Histidines and Charged Residues as Potential Triggers of a Conformational Change in the Fusion Loop of Ebola Virus Glycoprotein. PLoS One 11:e0152527
Yang, Sung-Tae; Lim, Sung In; Kiessling, Volker et al. (2016) Site-specific fluorescent labeling to visualize membrane translocation of a myristoyl switch protein. Sci Rep 6:32866
Yang, Sung-Tae; Kiessling, Volker; Simmons, James A et al. (2015) HIV gp41-mediated membrane fusion occurs at edges of cholesterol-rich lipid domains. Nat Chem Biol 11:424-31
Moissoglu, Konstadinos; Kiessling, Volker; Wan, Chen et al. (2014) Regulation of Rac1 translocation and activation by membrane domains and their boundaries. J Cell Sci 127:2565-76
Tamm, Lukas K; Lee, Jinwoo; Liang, Binyong (2014) Capturing glimpses of an elusive HIV gp41 prehairpin fusion intermediate. Structure 22:1225-1226

Showing the most recent 10 out of 44 publications