Telomerase from human leukemia cells : Properties and its interaction with deoxynucleoside analogues

Telomerase is a unique reverse transcriptase involved in the maintenance of genomic integrity. In an attempt to understand the properties of this enzyme and to study the effect of deoxynucleoside analogues, we have isolated and partially purified telomerase from the blast cells of a patient with acu...

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Bibliographic Details
Published in:Cancer research (Chicago, Ill.) Ill.), 1998-05, Vol.58 (9), p.1909-1913
Main Authors: PAI, R. B, PAI, S. B, KUKHANOVA, M, DUTSCHMAN, G. E, XIN GUO, CHENG, Y.-C
Format: Article
Language:English
Subjects:
Biological and medical sciences
Chromatography, Ion Exchange
Deoxyribonucleotides - pharmacology
Dose-Response Relationship, Drug
Electrophoresis, Polyacrylamide Gel
Endopeptidase K - pharmacology
Enzyme-Linked Immunosorbent Assay
General aspects (metabolism, cell proliferation, established cell line...)
Humans
Leukemia, Myeloid, Acute - enzymology
Medical sciences
Polymerase Chain Reaction
Potassium Chloride - pharmacology
Ribonuclease, Pancreatic - pharmacology
Telomerase - drug effects
Telomerase - isolation & purification
Telomerase - metabolism
Tumor cell
Tumor Cells, Cultured - enzymology
Tumors
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Summary:Telomerase is a unique reverse transcriptase involved in the maintenance of genomic integrity. In an attempt to understand the properties of this enzyme and to study the effect of deoxynucleoside analogues, we have isolated and partially purified telomerase from the blast cells of a patient with acute myelogenous leukemia. During the course of purification of telomerase, three characteristic forms of this enzyme activity were separated. Two processive forms and one less processive form were noted. All forms of the enzyme activities could be abolished by RNase A and proteinase K treatments, implying that they are ribonucleoproteins. The major form of telomerase was characterized with respect to divalent ion requirements, effect of salt and nonionic detergents. The Km of deoxynucleoside triphosphates was determined with a modified telomerase repeat array protocol assay. Studies with deoxynucleoside analogues indicated that 3'-azido-3'deoxythymidine triphosphate is much more inhibitory than 2',3'-dideoxy 2',3'didehydrothymidine triphosphate, and the cytidine analogue ddCTP was not inhibitory. ddGTP was the most potent inhibitor among all dideoxynucleosides studied.
ISSN:0008-5472
1538-7445