Critical role of reverse transcriptase in the inhibitory mechanism of CNI-H0294 on HIV-1 nuclear translocation.

Abstract

HIV-1 replication requires the translocation of viral genome into the nucleus of a target cell. We recently reported the synthesis of an arylene bis(methyl ketone) compound (CNI-H0294) that inhibits nuclear targeting of the HIV-1 genome and thus HIV-1 replication in monocyte cultures. Here we demonstrate that CNI-H0294 inhibits nuclear targeting of HIV-1-derived preintegration complexes by inactivating the nuclear localization sequence of the HIV-1 matrix antigen in a reaction that absolutely requires reverse transcriptase. This drug/reverse transcriptase interaction defines the specificity of its antiviral effect and is most likely mediated by the pyrimidine side-chain of CNI-H0294. After binding to reverse transcriptase, the carbonyl groups of CNI-H0294 react with the nuclear localization sequence of matrix antigen and prevent its binding to karyopherin a, the cellular receptor for nuclear localization sequences that carries proteins into the nucleus. Our results provide a basis for the development of a novel class of compounds that inhibit nuclear translocation and that can, in principle, be modified to target specific infectious agents.

HIV-1 Vpr interacts with the nuclear transport pathway to promote macrophage infection

“…MA and IN bind to importin-␣ as NLS-bearing substrates (Gallay et al 1996(Gallay et al , 1997. It has been suggested recently that HIV-1 Vpr binds importin-␣ at a site distinct from the NLS-binding site (Popov et al 1996), and it was shown that HIV infection of macrophages by Vpr-containing virions is not inhibited by NLS peptide (Gallay et al 1996(Gallay et al , 1997. Therefore, unlike MA and IN, Vpr does not require the importin-␣ NLS-binding site for its transport function.…”

Therapeutic Targeting of Nuclear Protein Import in Pathological Cell Conditions

“…Nuclear translocation of HIV-1 preintegration complex (PIC) (a nucleic acid/protein complex) is essential for viral replication, because it allows the PIC to get into contact with the cellular chromatin (Haffar et al, 2005). Arylene bis (methylketone) small-molecularweight compounds, such as the agent CNI-H1194, show therapeutic promise by disrupting the formation of the PIC, which could lead to inefficient nuclear import (Dubrovsky et al, 1995;Popov et al, 1996;Gasiorowski and Dean, 2003). However, there is still no consensus about the mechanisms that govern nuclear import of the HIV-1 PIC (Nisole and Saïb, 2004;Haffar et al, 2005).…”

Characterization of Antiviral Activity of Benzamide Derivative AH0109 against HIV-1 Infection

“…Alternatively, AH0109 may simultaneously affect both reverse transcription and viral nuclear import. A previous study reported that an anti-HIV-1 compound, CNI-H0294, binds to reverse transcriptase, and this interaction is required for CNI-H0294 to subsequently target the nuclear import signal of the HIV-1 MA protein and inhibit virus nuclear translocation (31). The latter study also suggested that modifications in a CNI-H0294 side chain would allow this compound to inhibit both RT activity and viral PIC nuclear translocation.…”