Format

Send to

Choose Destination
J Biol Chem. 2015 Apr 17;290(16):10504-17. doi: 10.1074/jbc.M114.626903. Epub 2015 Feb 27.

Identification of a novel HIV-1 inhibitor targeting Vif-dependent degradation of human APOBEC3G protein.

Author information

1
From the Departments of Cancer Immunology and AIDS and Departments of Pathology and.
2
Department of Biology, College of the Holy Cross, Worcester, Massachusetts 01610.
3
Southern Research Institute High Throughput Screening Center, Birmingham, Alabama 35205, and.
4
From the Departments of Cancer Immunology and AIDS and.
5
Cancer Biology, Dana Farber Cancer Institute and.
6
Southern Research Institute, Department of Infectious Disease Research, Frederick, Maryland 21701.
7
From the Departments of Cancer Immunology and AIDS and Neurology (Microbiology), Harvard Medical School, Boston, Massachusetts 02115, dana_gabuzda@dfci.harvard.edu.

Abstract

APOBEC3G (A3G) is a cellular cytidine deaminase that restricts HIV-1 replication by inducing G-to-A hypermutation in viral DNA and by deamination-independent mechanisms. HIV-1 Vif binds to A3G, resulting in its degradation via the 26 S proteasome. Therefore, this interaction represents a potential therapeutic target. To identify compounds that inhibit interaction between A3G and HIV-1 Vif in a high throughput format, we developed a homogeneous time-resolved fluorescence resonance energy transfer assay. A 307,520 compound library from the NIH Molecular Libraries Small Molecule Repository was screened. Secondary screens to evaluate dose-response performance and off-target effects, cell-based assays to identify compounds that attenuate Vif-dependent degradation of A3G, and assays testing antiviral activity in peripheral blood mononuclear cells and T cells were employed. One compound, N.41, showed potent antiviral activity in A3G(+) but not in A3G(-) T cells and had an IC50 as low as 8.4 μM and a TC50 of >100 μM when tested against HIV-1Ba-L replication in peripheral blood mononuclear cells. N.41 inhibited the Vif-A3G interaction and increased cellular A3G levels and incorporation of A3G into virions, thereby attenuating virus infectivity in a Vif-dependent manner. N.41 activity was also species- and Vif-dependent. Preliminary structure-activity relationship studies suggest that a hydroxyl moiety located at a phenylamino group is critical for N.41 anti-HIV activity and identified N.41 analogs with better potency (IC50 as low as 4.2 μM). These findings identify a new lead compound that attenuates HIV replication by liberating A3G from Vif regulation and increasing its innate antiviral activity.

KEYWORDS:

Antiviral Agent; Cytidine Deaminase; High Throughput Screening (HTS); Human Immunodeficiency Virus (HIV); Viral Protein; Virology

PMID:
25724652
PMCID:
PMC4400358
DOI:
10.1074/jbc.M114.626903
[Indexed for MEDLINE]
Free PMC Article

Publication type, MeSH terms, Substances, Grant support

Publication type

MeSH terms

Substances

Grant support

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center