Identification of a key target sequence to block human immunodeficiency virus type 1 replication within the gag-pol transframe domain.
Although the full sequence of the human immunodeficiency virus type 1 (HIV-1) genome has been known for more than a decade, effective genetic antivirals have yet to be developed. Here we show that, of 22 regions examined, one highly conserved sequence (ACTCTTTGGCAACGA) near the 3' end of the HIV-1 gag-pol transframe region, encoding viral protease residues 4 to 8 and a C-terminal Vpr-binding motif of p6(Gag) protein in two different reading frames, can be successfully targeted by an antisense peptide nucleic acid oligomer named PNA(PR2). A disrupted translation of gag-pol mRNA induced at the PNA(PR2)-annealing site resulted in a decreased synthesis of Pr160(Gag-Pol) polyprotein, hence the viral protease, a predominant expression of Pr55(Gag) devoid of a fully functional p6(Gag) protein, and the excessive intracellular cleavage of Gag precursor proteins, hindering the processes of virion assembly. Treatment with PNA(PR2) abolished virion production by up to 99% in chronically HIV-1-infected H9 cells and in peripheral blood mononuclear cells infected with clinical HIV-1 isolates with the multidrug-resistant phenotype. This particular segment of the gag-pol transframe gene appears to offer a distinctive advantage over other regions in invading viral structural genes and restraining HIV-1 replication in infected cells and may potentially be exploited as a novel antiviral genetic target.
Sei
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HIV Clinical Interface Laboratory, SAIC-Frederick, NCI-Frederick Cancer Research and Development Center, Frederick, Maryland 21702, USA. SEI@dtpax2.ncifcrf.gov
Yang
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O'Neill
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Yoshimura
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Nagashima
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Mitsuya
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D020413:3' Untranslated Regions / Q000235:genetics; D019380:Anti-HIV Agents / Q000494:pharmacology*; D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D004789:Enzyme Activation; D015685:Fusion Proteins, gag-pol / Q000235:genetics* / Q000378:metabolism; D015683:Gene Products, gag / Q000096:biosynthesis / Q000378:metabolism; D018390:Gene Targeting; D016655:HIV Core Protein p24 / Q000096:biosynthesis; D015658:HIV Infections / Q000821:virology; D016333:HIV Protease / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011498:Protein Precursors / Q000096:biosynthesis / Q000378:metabolism; D014771:Virion / Q000502:physiology; D014779:Virus Replication / Q000187:drug effects*; D054301:gag Gene Products, Human Immunodeficiency Virus; D054302:pol Gene Products, Human Immunodeficiency Virus
D020413:3' Untranslated Regions; D019380:Anti-HIV Agents; D015685:Fusion Proteins, gag-pol; D015683:Gene Products, gag; D016655:HIV Core Protein p24; D020135:Peptide Nucleic Acids; D011498:Protein Precursors; D054301:gag Gene Products, Human Immunodeficiency Virus; C059767:p55 gag precursor protein, Human immunodeficiency virus 1; D054302:pol Gene Products, Human Immunodeficiency Virus; C091950:pr160 gag-pol precursor protein, Human immunodeficiency virus 1; D016333:HIV Protease