Detection of telomerase activity in Chinese hamster V79 cells and its inhibition by 7-deaza-deoxy guanosine triphosphate and (TTAGGG)4 in vitro.
To investigate the nature of telomerase activity and its inhibition in Chinese hamster V79 cells, we have detected telomerase activity in Chinese hamster cells using Telomeric Repeat Amplification Protocol (TRAP) assay. We have further studied inhibition characteristics of this enzyme in vitro by nucleotide analogue 7-deaza-2'-deoxy guanosine triphosphate (7-deaza-dGTP) and oligonucleotide (TTAGGG)4. Both the inhibitors inhibited the telomerase activity in a dose dependent manner. To attain 50% inhibition of the telomerase activity, we needed about 4.5 microM of 7-deaza-dGTP. Similarly, preincubation at 37 degreesC of the cell extract with 1.25 x 10(-3) microgram oligonucleotide (TTAGGG)4 showed 50% inhibition of the control value. Inhibition of telomerase activity by 7-deaza-dGTP could be due to the incorporation of the modified nucleotide in the telomeric repeat and thus altering the further binding/extension by the enzyme. (TTAGGG)4 could have possibly interacted with RNA component of telomerase and inhibited its activity.
Pandit
B
B
Crystallography and Molecular Biology Division, Saha Institute of Nuclear Physics, Calcutta-, 1/AF Bidhan Nagar, 700 064, India.
Bhattacharyya
N P
NP
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D007700:Kinetics; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D019098:Telomerase / Q000037:antagonists & inhibitors / Q000378:metabolism*
D009838:Oligodeoxyribonucleotides; C045550:7-deaza-2'-deoxyguanosine; D019098:Telomerase; D003849:Deoxyguanosine