Targeting HIV-1 integrase with aptamers selected against the purified RNase H domain of HIV-1 RT.
Several in vitro strategies have been developed to selectively screen for nucleic acid sequences that bind to specific proteins. We previously used the SELEX procedure to search for aptamers against HIV-1 RNase H activity associated with reverse transcriptase (RT) and human RNase H1. Aptamers containing G-rich sequences were selected in both cases. To investigate whether the interaction with G-rich oligonucleotides (ODNs) was a characteristic of these enzymes, a second in vitro selection was performed with an isolated RNase H domain of HIV-1 RT (p15) as a target and a new DNA library. In this work we found that the second SELEX led again to the isolation of G-rich aptamers. But in contrast to the first selection, these latter ODNs were not able to inhibit the RNase H activity of either the p15 domain or the RNase H embedded in the complete RT. On the other hand, the aptamers from the first SELEX that were inhibitors of the RT-associated RNase H did not inhibit the activity of the isolated p15 domain. This suggests that the active conformation of both RNase H domains is different according to the presence or absence of the DNA polymerase domain. HIV-1 RNase H and integrase both belong to the phosphotransferase family and share structural similarities. An interesting result was obtained when the DNA aptamers initially raised against p15 RNase H were assayed against HIV-1 integrase. In contrast to RNase H, the HIV-1 integrase was inhibited by these aptamers. Our results point out that prototype structures can be exploited to develop inhibitors of two related enzymes.
Métifiot
Mathieu
M
UMR 5097 CNRS, Université Victor Segalen Bordeaux 2, 146, rue Léo Saignat, 33076 Bordeaux cedex, France.
Leon
Oscar
O
Tarrago-Litvak
Laura
L
Litvak
Simon
S
Andréola
Marie-Line
ML
D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000494:pharmacology*; D001483:Base Sequence; D019427:HIV Integrase / Q000737:chemistry / Q000187:drug effects* / Q000302:isolation & purification; D019428:HIV Integrase Inhibitors / Q000494:pharmacology*; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000201:enzymology*; D006801:Humans; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism*; D016914:Ribonuclease H / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D013698:Templates, Genetic
D052157:Aptamers, Nucleotide; D019428:HIV Integrase Inhibitors; D009841:Oligonucleotides; D019427:HIV Integrase; D016914:Ribonuclease H