This Project is designed to characterize the early post-entry steps of HIV-1 replication, core evolution, and TRIM5? restriction in structural and mechanistic detail. Following virus-cell fusion, the viral genomic RNA (gRNA) is reverse transcribed1-6, the core interacts with host factors and loses its capsid (?uncoating?)7-11, and the host restriction factor TRIM5? can recognize the capsid and block these transformations.12-16 To characterize these processes, we will determine 3D structures of replication initiation complexes that comprise reverse transcriptase (RT), the host tRNALys,3 primer, and various gRNA templates. We will study the conformational changes that accompany replication (Aim 1), develop and apply new single particle assays for studying reverse transcription within isolated viral cores (Aim 2), image transduced cells using fluorescent microscopy and correlated light and electron cryotomography (CLEM-ECT) to visualize and characterize core trafficking, reverse transcription, uncoating and restriction (Aim 3), and reconstitute, image and model TRIM5?- capsid complexes to learn how TRIM5? inactivates the core and stimulates innate immune responses (Aim 4).

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Specialized Center (P50)
Project #
2P50GM082545-11
Application #
9411510
Study Section
Special Emphasis Panel (ZRG1)
Project Start
Project End
Budget Start
2017-08-01
Budget End
2018-07-31
Support Year
11
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Utah
Department
Type
DUNS #
009095365
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
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Bailey, Lucas J; Sheehy, Kimberly M; Dominik, Pawel K et al. (2018) Locking the Elbow: Improved Antibody Fab Fragments as Chaperones for Structure Determination. J Mol Biol 430:337-347

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