User:Nathan Roy
User:Nathan Roy/Sandbox 1I am a graduate student at the University of Vermont, studying the dynamics of HIV-1 cell to cell transmission and HIV-1 induced syncytia formation. Our wonderful professor, Dr. Steven Everse, has commissioned us (his students from his BioChem 351 course) to create a page describing the structure of a protein that interests us. I have chosen the HIV-1 gag protein, and more specifically, the MA and CA domains.
The HIV-1 Gag protein is the major structural protein required for virus assembly. It is synthesized as a polyprotein in the cytosol of an infected cell, and contains four functional segments; MA, CA (NTD and CTD), NC, and p6. The NC region is flanked by two "spacer" segments, denoted SP1 and SP2. The polyprotein is all alpha helical, except the NC region, which is composed RNA interacting beta sheets. Gag is often referred to as an "assembly machine", because expression of Gag alone is sufficient to produce budding virus-like particles (VLP's). Here, we will take a closer look at the MA and CA domains, and how the structural components of these domains aid in the assembly of virus particles.
The MA domain (also called Matrix) is essential for proper targeting of Gag (and thus virus release) to distinct locations on the plasma membrane. The MA domain is myristylated post translationally, which is important for a stable association with the plasma membrane. Perhaps just as important for proper virus assembly, is the interaction of MA with phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2). It has long been known that HIV-1 budding occurs at sites rich in PI(4,5)P2 and cholesterol (lipid rafts, and tetraspanin enriched microdomains, TEMs), probably to ensure proper virus budding localized to the plasma membrane (elimination of PI(4,5)P2 causes virions to assembly at intracellular endosomes). More recently, structural analysis of MA has revealed a myristylation switch that allows exposer of the myristyl group to the plasma membrane only upon binding of MA to PI(4,5)P2, a mechanism ensuring proper localization of Gag to rafts within the plasma membrane.
When MA is not bound to PI(4,5)P2 (Figure 1)
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, notice the alignment of helix 1, and more precisely, the orientation of Leu 8 and Glu 12. INSERT SCENE SHOWING RESIDUES In this PI(4,5)P2 unbound structure, the myristyl group is sequestered in the pocket of helix 1 created by Leu 8 and Glu 12. Upon binding of PI(4,5)P2 to the hydrophobic groove created by helix 2, a type 2 beta turn, and helix 5, a slight conformational switch occurs in helix 1 (Figure 2),
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causing a change in the alignment of Leu 8 and Glu 12, ejecting the myristyl group from it's sequestered state. INSERT SCENE SHOWING RESIDUES