Viruses invade all forms of life, causing diseases in humans including HIV-AIDS. Although the variety of viruses is daunting, all enveloped viruses, including HIV-1 and Rous Sarcoma Virus (RSV), must associate with the inner leaflet of plasma membranes. Our laboratories have undertaken a large-scale effort to characterize interactions between membrane components. The goal is to discover which membrane factors exert control over the interactions between membrane-associated viral matrix proteins. The overall approach is to combine experimental measurements, mainly FRET and fluorescence microscopy, with Monte Carlo computer simulations in order to discover all of the significant interaction energies between lipid-lipid, lipid-protein, and protein-protein. Membrane phase behavior and protein-protein interactions are first measured in model liposomes and then in live chicken fibroblast cells. The first specific aim is to develop the experiments and Monte Carlo simulations to describe gramicidin-gramicidin interactions in a simple 3-lipid component mixture containing cholesterol.
The second aim i s to handle the greatly increased complexity of a fifth component, ceramide or diacylglycerol.
This aim i ncludes proposed development of a """"""""lipid buffer"""""""" in order to control the chemical potential of cholesterol, ceramide, or diacylglycerol in model or real biomembranes.
The third aim i s to characterize HIV-1 and RSV matrix protein-protein interactions in a lipid mixture that models the inner leaflet of an animal cell plasma membrane, as a basis for understanding behavior in live cells. The fourth and final aim is to examine HIV-1 and RSV matrix protein-protein interactions at the inner leaflet of the plasma membrane using FRET in live cells, attempting to find the factors that control the interactions, and whether coupling to the outer leaflet is observed. Useful treatments for viral afflictions, in particular HIV-AIDS, are not limited to """"""""all or none"""""""" cures. Combinations of drugs, each of which slows down a step of the viral life cycle, have proven useful. The proposed research will lead to detailed understanding of how the membrane components cholesterol, ceramide, and diacylglycerol, all of which can be partially regulated by drugs, control one critical step in the life of the virus - association/assembly at the plasma membrane.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM077198-04
Application #
7776840
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Chin, Jean
Project Start
2007-03-10
Project End
2012-02-28
Budget Start
2010-03-01
Budget End
2012-02-28
Support Year
4
Fiscal Year
2010
Total Cost
$367,960
Indirect Cost
Name
Cornell University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
872612445
City
Ithaca
State
NY
Country
United States
Zip Code
14850
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Alwarawrah, Mohammad; Dai, Jian; Huang, Juyang (2012) Modification of Lipid Bilayer Structure by Diacylglycerol: A Comparative Study of Diacylglycerol and Cholesterol. J Chem Theory Comput 8:749-758
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Dai, Jian; Alwarawrah, Mohammad; Ali, Md Rejwan et al. (2011) Simulation of the lo-ld phase boundary in DSPC/DOPC/cholesterol ternary mixtures using pairwise interactions. J Phys Chem B 115:1662-71
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Morales-Penningston, Nelson F; Wu, Jing; Farkas, Elaine R et al. (2010) GUV preparation and imaging: minimizing artifacts. Biochim Biophys Acta 1798:1324-32
Alwarawrah, Mohammad; Dai, Jian; Huang, Juyang (2010) A molecular view of the cholesterol condensing effect in DOPC lipid bilayers. J Phys Chem B 114:7516-23

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