Sequence-specific cleavage activities of DNA enzymes targeted against HIV-1 Gag and Nef regions.
The genome of HIV-1 is known to accumulate nucleotide changes throughout the course of disease that result in generation of escape mutants. Therefore, any nucleic acid-based antiviral approach should be targeted against multiple regions of the HIV-1 genome that might significantly delay the appearance of such mutants. We designed several DNA enzymes against the most conserved p24 Gag and the Nef regions in the HIV-1 genome. Sequence-specific cleavage activity was observed for all the DNA enzymes tested. Gag DNA enzyme, which cleaved the target RNA more efficiently in the presence of low levels or physiologic levels of Mg(2+), interfered more effectively with HIV-1 gene expression in virus challenge experiments. The two Nef DNA enzymes, as observed with Gag DNA enzymes, showed significant variation in their cleavage activities in the presence of varying concentration of Mg(2+) and, as expected, did not interfere with the replication of a laboratory-adapted HIV-1 isolate under in vitro culture conditions. The Gag DNA enzymes could be exploited in combination with other promising antiviral approaches.
Dash
Bipin C
BC
Laboratory of Virology, National Institute of Immunology, JNU Campus, New Delhi-110067, India.
Banerjea
Akhil C
AC
D001483:Base Sequence; D017931:DNA Primers; D021881:DNA, Catalytic / Q000378:metabolism*; D015683:Gene Products, gag / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015693:Gene Products, nef / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016679:Genome, Viral; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D006868:Hydrolysis; D015636:Magnesium Chloride / Q000737:chemistry; D014779:Virus Replication; D054311:nef Gene Products, Human Immunodeficiency Virus
D017931:DNA Primers; D021881:DNA, Catalytic; D015683:Gene Products, gag; D015693:Gene Products, nef; D054311:nef Gene Products, Human Immunodeficiency Virus; D015636:Magnesium Chloride