Interaction of aurintricarboxylic acid (ATA) with four nucleic acid binding proteins DNase I, RNase A, reverse transcriptase and Taq polymerase

Abstract

In the investigation of interaction of aurintricarboxylic acid (ATA) with four biologically important proteins we observed inhibition of enzymatic activity of DNase I, RNase A, M-MLV reverse transcriptase and Taq polymerase by ATA in vitro assay. As the telomerase reverse transcriptase (TERT) is the main catalytic subunit of telomerase holoenzyme, we also monitored effect of ATA on telomerase activity in vivo and observed dose-dependent inhibition of telomerase activity in Chinese hamster V79 cells treated with ATA. Direct association of ATA with DNase I (K(d)=9.019 microM)), RNase A (K(d)=2.33 microM) reverse transcriptase (K(d)=0.255 microM) and Taq polymerase (K(d)=81.97 microM) was further shown by tryptophan fluorescence quenching studies. Such association altered the three-dimensional conformation of DNase I, RNase A and Taq polymerase as detected by circular dichroism. We propose ATA inhibits enzymatic activity of the four proteins through interfering with DNA or RNA binding to the respective proteins either competitively or allosterically, i.e. by perturbing three-dimensional structure of enzymes.

dsRNA binding characterization of full length recombinant wild type and mutants Zaire ebolavirus VP35

“…Therefore, we wanted to validate our newly established magnetic pull down assay also as a method for the screening of molecules that may inhibit the interaction between dsRNA and VP35. In the absence of a specific VP35-dsRNA binding inhibitor, we used a known inhibitor of nucleic acid-protein interactions such as the ATA (Ghosh et al, 2009;Gonzáles et al, 1980) (Fig. 5).…”

Aurintricarboxylic Acid Modulates the Affinity of Hepatitis C Virus NS3 Helicase for Both Nucleic Acid and ATP

“…2, 3 The molecular basis for the sensitivity of nucleic acid enzymes to submicromolar ATA concentrations likely results from the ability of ATA to form polymers in solution that resemble nucleic acids (Fig. 1).…”

“…4, 5 ATA polymers bind RNA 6 and also bind to nucleic acid binding clefts on nucleases and similar proteins. 3, 4, 7–10 …”

Suppressing RNA silencing with small molecules and the viral suppressor of RNA silencing protein p19

“…Our results indicate that ATA is a potent inhibitor of p19:siRNA interactions, inhibiting p19 with the same potency as observed for AGO-2 (Table 1) [32]. This suggests that p19 and ATA are not compatible approaches for inhibiting the RNA silencing pathway in biological settings [33].…”

“…ATA is a triphenylmethane molecule which inhibits a wide range of nucleic acid binding proteins [33,34]. Oxidopamine is an adrenergic compound applied in research to induce symptoms of Parkinson's disease in animal models [35].…”