The objective of the proposed work is to determine the mechanism of particle maturation for the HK97 virus, a highly accessible model system. Virtually all complex viruses initially assemble into non-infectious, often fragile, particles that are called provirions. Encoded in their structure is a program that directs particle conformational changes leading to a robust, infectious virion. Six different intermediates of this maturation have been isolated for HK97. Previous support for this work lead to the near-atomic resolution structures of three intermediates and the discovery that early maturation is an exothermic process and later maturation is driven by a Brownian ratchet mechanism. We now propose to determine the structures of three more intermediates and to study the transitions between all of the intermediates with single particle and ensemble methods. Our goal is to understand how the expression of two viral gene products creates a meta-stable particle that releases ~20kCal/mole of energy in the initial step of maturation and then proceeds by biasing thermal fluctuations with a covalent cross-link to complete the maturation process. We are also investigating the use of the mature particles as a potential delivery system for drugs in nano medicine.

Public Health Relevance

Project Narrative Virus maturation is virtually universal among complex viruses and is a worthy target for antiviral therapy. It is, however, very difficult to study because purified virions are fully mature. Grasping the details of the process requires accessible model systems where intermediates can be isolated and studied, as well as the transitions between them. HK97 is such a system and we are exploiting it to understand the structural, kinetic and thermodynamic parameters of particle maturation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI040101-17
Application #
8239574
Study Section
Special Emphasis Panel (ZRG1-BCMB-B (03))
Program Officer
Park, Eun-Chung
Project Start
1996-07-01
Project End
2015-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
17
Fiscal Year
2012
Total Cost
$423,002
Indirect Cost
$200,252
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Lakritz, Jessica R; Thibault, Derek M; Robinson, Jake A et al. (2016) α4-Integrin Antibody Treatment Blocks Monocyte/Macrophage Traffic to, Vascular Cell Adhesion Molecule-1 Expression in, and Pathology of the Dorsal Root Ganglia in an SIV Macaque Model of HIV-Peripheral Neuropathy. Am J Pathol 186:1754-61
Lakritz, Jessica R; Robinson, Jake A; Polydefkis, Michael J et al. (2015) Loss of intraepidermal nerve fiber density during SIV peripheral neuropathy is mediated by monocyte activation and elevated monocyte chemotactic proteins. J Neuroinflammation 12:237
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Lakritz, Jessica R; Bodair, Ayman; Shah, Neal et al. (2015) Monocyte Traffic, Dorsal Root Ganglion Histopathology, and Loss of Intraepidermal Nerve Fiber Density in SIV Peripheral Neuropathy. Am J Pathol 185:1912-23
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