High affinity interaction of HIV‐1 integrase with specific and non‐specific single‐stranded short oligonucleotides

Abstract

Retroviral integrase (IN) catalyzes the integration of double-stranded viral DNA into the host cell genome. The reaction can be divided in two steps: 3P P-end processing and DNA strand transfer. Here we studied the effect of short oligonucleotides (ODNs) on human immunodeficiency virus type 1 (HIV-1) IN. ODNs were either specific, with sequences representing the extreme termini of the viral long terminal repeats, or nonspecific. All ODNs were found to competitively inhibit the processing reaction with K i values in the nM range for the best inhibitors. Our studies on the interaction of IN with ODNs also showed that: (i) besides the 3P P-terminal GT, the interaction of IN with the remaining nucleotides of the 21-mer specific sequence was also important for an effective interaction of the enzyme with the substrate; (ii) in the presence of specific ODNs the activity of the enzyme was enhanced, a result which suggests an ODNinduced conformational change of HIV-1 IN.z 1999 Federation of European Biochemical Societies.

The lethal phenotype observed after HIV-1 integrase expression in yeast cells is related to DNA repair and recombination events

“…Purification was performed essentially as previously described (Caumont et al, 1999). The soluble fraction containing the HIV-1 IN obtained from JSC 310 (IN) expressing IN protein was loaded on a Hitrap butyl-sepharose 4B column (1 ml, Pharmacia-LKB), washed with LSC buffer (50 mM HEPES pH 7.6, 0.2 M NaCl, 0.1 M EDTA, 1 mM DTT, 7 mM CHAPS, 10% glycerol) and equilibrated with 5-volume HSC buffer (50 mM HEPES pH 7.6, 0.2 M NaCl, 1 M ammonium sulfate, 0.1 mM EDTA, 1 mM DTT, 7 mM CHAPS).…”

“…Recombinant HIV-1 IN was obtained from the JSC310 yeast strain transformed with the integrase expression plasmid, pHIV1SF2IN, in culture conditions allowing a high level of IN expression (Caumont et al, 1999). Both wildtype and mutated D116A integrases were purified and assayed for processing.…”

Functional Interactions of Human Immunodeficiency Virus Type 1 Integrase with Human and Yeast HSP60

“…Purification was performed essentially as previously described (6). The soluble fraction containing the HIV-1 IN obtained from JSC 310 (IN) was loaded on a Hitrap butylSepharose 4B column (1 ml; Pharmacia-LKB), washed with LSC buffer (50 mM HEPES, pH 7.6; 0.2 M NaCl; 0.1 M EDTA; 1 mM DTT; 7 mM CHAPS; 10% glycerol) and equilibrated with 5 volumes of HSC buffer (50 mM HEPES, pH 7.6; 0.2 M NaCl; 1 M ammonium sulfate; 0.1 mM EDTA; 1 mM DTT; 7 mM CHAPS).…”

The inhibition process of HIV-1 integrase by diketoacids molecules: Understanding the factors governing the better efficiency of dolutegravir

“…These reveal strong non-covalent interactions within the drug-DNA complex. In addition, the green plot represented in Figure 4b for the DTG complex has broader extensions than the one corresponding to the EVG complex (Figure 4a [7,20,28,[33][34][35].…”

“…There appear to be stringent requirements concerning the sequence of the nucleic bases at the ends of the LTR for retroviral integration [35][36][37][38][39][40], and there is no evidence of mutations in …”