Human AP0BEC3G (A3G ) mutates the HIV genome by deaminafing cDNA cytosines to uracils. This activity can inactivate HIV. However, as a counter-defense, HIV uses an auxiliary protein called Vif (virion infectivity factor) to degrade A3G. A direct protein-protein interaction is required for Vif to neutralize A3G. Although, high-resolution structures of the deaminase domain of A3G have been achieved recently (including two NMR structures from Matsuo laboratory), structures of the full-length A3G protein and its complexes with DNA or Vif have proven more elusive. APOBEC3F (A3F) is another potent HIV-1 restriction factor. Much less structural information is available for A3F as there is no high-resolution structure for this protein. Structural and biochemical studies of A3G and A3F have been hinderd by their insolubility. We have recently generated A3G and A3F variants that are folded, catalytically acitive and soluble enough to take NMR spectra. We propose to solve structures of full-length A3G, the catalytic domain of A3F and their complexes with Vif (or Vif fragments) using NMR. We will also interrogate the interactions between A3G/F and single-stranded DNA. Our objectives are to reveal structurally (1) intra- and inter-molecular domain-domain interaction of A3G and A3F, (2) their mechanisms for binding ssDNA and (S) similarities and differences between A3G and A3F in their Vif-binding domains. These studies are an integral part of a larger interdisciplinary program project that will generate unprecedented molecular, biochemical, biophysical and structural information on this critical host-pathogen interaction.

Public Health Relevance

Structural information of the HIV-1 restriction factors A3G and A3F will provide a strong foundation for further biochemical and cellular studies. In addition, an atomic-level understanding of the A3-Vif interaction can be used to rationally develop new therapeutic methods for HIV/AIDS.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
5P01GM091743-02
Application #
8375090
Study Section
Special Emphasis Panel (ZRG1-AARR-D)
Project Start
Project End
Budget Start
2012-01-01
Budget End
2012-12-31
Support Year
2
Fiscal Year
2012
Total Cost
$292,634
Indirect Cost
$64,322
Name
University of Minnesota Twin Cities
Department
Type
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Richards, Christopher M; Li, Ming; Perkins, Angela L et al. (2017) Reassessing APOBEC3G Inhibition by HIV-1 Vif-Derived Peptides. J Mol Biol 429:88-96
Pan, Yangang; Sun, Zhiqiang; Maiti, Atanu et al. (2017) Nanoscale Characterization of Interaction of APOBEC3G with RNA. Biochemistry 56:1473-1481
Kouno, Takahide; Silvas, Tania V; Hilbert, Brendan J et al. (2017) Crystal structure of APOBEC3A bound to single-stranded DNA reveals structural basis for cytidine deamination and specificity. Nat Commun 8:15024
Khisamutdinov, Emil F; Jasinski, Daniel L; Li, Hui et al. (2016) Fabrication of RNA 3D Nanoprisms for Loading and Protection of Small RNAs and Model Drugs. Adv Mater 28:10079-10087
Shlyakhtenko, Luda S; Dutta, Samrat; Li, Ming et al. (2016) Single-Molecule Force Spectroscopy Studies of APOBEC3A-Single-Stranded DNA Complexes. Biochemistry 55:3102-6
Li, Jinhui; Barylko, Barbara; Eichorst, John P et al. (2016) Association of Endophilin B1 with Cytoplasmic Vesicles. Biophys J 111:565-576
Sharma, Ashwani; Haque, Farzin; Pi, Fengmei et al. (2016) Controllable self-assembly of RNA dendrimers. Nanomedicine 12:835-844
Shaban, Nadine M; Shi, Ke; Li, Ming et al. (2016) 1.92 Angstrom Zinc-Free APOBEC3F Catalytic Domain Crystal Structure. J Mol Biol 428:2307-16
Lyubchenko, Yuri L; Shlyakhtenko, Luda S (2016) Imaging of DNA and Protein-DNA Complexes with Atomic Force Microscopy. Crit Rev Eukaryot Gene Expr 26:63-96
Li, Hui; Zhang, Kaiming; Pi, Fengmei et al. (2016) Controllable Self-Assembly of RNA Tetrahedrons with Precise Shape and Size for Cancer Targeting. Adv Mater 28:7501-7

Showing the most recent 10 out of 82 publications