The major goal of this competing renewal is to define viral interactions with host pathways that are essential for the assembly of infectious HIV-1 particles. HIV-1 particle assembly occurs on the plasma membrane of infected cells. A major finding from this project was the identification of Rab11-FIP1C (FIP1C) as a key adaptor protein required for HIV-1 envelope protein (Env) trafficking and particle incorporation. FIP1C-dependent trafficking underlies the cell type-specific incorporation of Env, as disruption of a key motif in the cytoplasmic tail (CT) involved in FIP1C translocation led to a loss of Env incorporation in cell types that was nearly identical to that of CT-deleted viruses. Our recent data establish that Env traverses the endosomal recycling compartment, where it meets FIP1C in a step required for subsequent outward trafficking and particle incorporation of relevant primary isolate Envs. Simian Immunodeficiency Virus (SIV) Envs and a subset of HIV Envs, in contrast, do not follow FIP1C-dependent trafficking in human cells. This competing renewal will fully define the viral and host determinants of Env trafficking pathways. Experiments here will identify additional members of the HIV-1 Env trafficking complex, including the essential kinesin mediating outward sorting of Env. Experiments in this application will define the sequential steps in Env trafficking that lead to the production of infectious viral particles, using a novel imaging strategy. FIP1C-mediated trafficking will be characterized in primary monocyte-derived macrophages, where membrane dynamics and the intracellular virus-containing compartment may introduce unique features. The essential role of FIP1C in the formation of the virological synapse and in macrophage-to-T cell transmission events will then be delineated.

Public Health Relevance

This project will define important pathways that allow infectious HIV particles to assemble. Experiments here will identify cellular components of the cellular pathway that is used by the HIV envelope protein to gain access to budding particles. Assembly of HIV particles, including envelope protein incorporation, is an understudied area of HIV biology that is relevant for what it can teach us about virus-host interactions and potentially can reveal new targets for antiviral therapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM111027-22A1
Application #
9694826
Study Section
AIDS Molecular and Cellular Biology Study Section (AMCB)
Program Officer
Sakalian, Michael
Project Start
1997-04-01
Project End
2023-01-31
Budget Start
2019-02-01
Budget End
2020-01-31
Support Year
22
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Cincinnati Children's Hospital Medical Center
Department
Type
DUNS #
071284913
City
Cincinnati
State
OH
Country
United States
Zip Code
45229
Spearman, Paul (2018) Viral interactions with host cell Rab GTPases. Small GTPases 9:192-201
Kirschman, Junghwa; Qi, Mingli; Ding, Lingmei et al. (2018) HIV-1 Envelope Glycoprotein Trafficking through the Endosomal Recycling Compartment Is Required for Particle Incorporation. J Virol 92:
Staitieh, Bashar S; Ding, Lingmei; Neveu, Wendy A et al. (2017) HIV-1 decreases Nrf2/ARE activity and phagocytic function in alveolar macrophages. J Leukoc Biol 102:517-525
Hammonds, Jason E; Beeman, Neal; Ding, Lingmei et al. (2017) Siglec-1 initiates formation of the virus-containing compartment and enhances macrophage-to-T cell transmission of HIV-1. PLoS Pathog 13:e1006181
Hampton, Cheri M; Strauss, Joshua D; Ke, Zunlong et al. (2017) Correlated fluorescence microscopy and cryo-electron tomography of virus-infected or transfected mammalian cells. Nat Protoc 12:150-167
Spearman, Paul (2016) HIV-1 Gag as an Antiviral Target: Development of Assembly and Maturation Inhibitors. Curr Top Med Chem 16:1154-66
St Gelais, Corine; Kim, Sun Hee; Ding, Lingmei et al. (2016) A Putative Cyclin-binding Motif in Human SAMHD1 Contributes to Protein Phosphorylation, Localization, and Stability. J Biol Chem 291:26332-26342
Chukwuma, Valentine U; Hicar, Mark D; Chen, Xuemin et al. (2015) Association of VH4-59 Antibody Variable Gene Usage with Recognition of an Immunodominant Epitope on the HIV-1 Gag Protein. PLoS One 10:e0133509
Qi, Mingli; Chu, Hin; Chen, Xuemin et al. (2015) A tyrosine-based motif in the HIV-1 envelope glycoprotein tail mediates cell-type- and Rab11-FIP1C-dependent incorporation into virions. Proc Natl Acad Sci U S A 112:7575-80
Giroud, Charline; Marin, Mariana; Hammonds, Jason et al. (2015) P2X1 Receptor Antagonists Inhibit HIV-1 Fusion by Blocking Virus-Coreceptor Interactions. J Virol 89:9368-82

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