Interaction of aurintricarboxylic acid (ATA) with four nucleic acid binding proteins DNase I, RNase A, reverse transcriptase and Taq polymerase.
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.
Ghosh
Utpal
U
Department of Biochemistry & Biophysics, University of Kalyani, Kalyani 741235, India. utpal8ghosh@gmail.com
Giri
Kalyan
K
Bhattacharyya
Nitai P
NP
D000818:Animals; D001312:Aurintricarboxylic Acid / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D002460:Cell Line; D002942:Circular Dichroism; D003850:Deoxyribonuclease I / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006041:Goats; D007700:Kinetics; D016133:Polymerase Chain Reaction; D012313:RNA / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D012259:Ribonuclease, Pancreatic / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D019914:Taq Polymerase / Q000378:metabolism*; D019098:Telomerase / Q000037:antagonists & inhibitors; D013696:Temperature
D001312:Aurintricarboxylic Acid; D012313:RNA; D019914:Taq Polymerase; D012194:RNA-Directed DNA Polymerase; D019098:Telomerase; D003850:Deoxyribonuclease I; D012259:Ribonuclease, Pancreatic