Effects of reverse transcriptase inhibitors on telomere length and telomerase activity in two immortalized human cell lines.
The ribonucleoprotein telomerase, a specialized cellular reverse transcriptase, synthesizes one strand of the telomeric DNA of eukaryotes. We analyzed telomere maintenance in two immortalized human cell lines: the B-cell line JY616 and the T-cell line Jurkat E6-1, and determined whether known inhibitors of retroviral reverse transcriptases could perturb telomere lengths and growth rates of these cells in culture. Dideoxyguanosine (ddG) caused reproducible, progressive telomere shortening over several weeks of passaging, after which the telomeres stabilized and remained short. However, the prolonged passaging in ddG caused no observable effects on cell population doubling rates or morphology. Azidothymidine (AZT) caused progressive telomere shortening in some but not all T- and B-cell cultures. Telomerase activity was present in both cell lines and was inhibited in vitro by ddGTP and AZT triphosphate. Prolonged passaging in arabinofuranyl-guanosine, dideoxyinosine (ddI), dideoxyadenosine (ddA), didehydrothymidine (d4T), or phosphonoformic acid (foscarnet) did not cause reproducible telomere shortening or decreased cell growth rates or viabilities. Combining AZT, foscarnet, and/or arabinofuranyl-guanosine with ddG did not significantly augment the effects of ddG alone. Strikingly, with or without inhibitors, telomere lengths were often highly unstable in both cell lines and varied between parallel cell cultures. We propose that telomere lengths in these T- and B-cell lines are determined by both telomerase and telomerase-independent mechanisms.
Strahl
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Department of Microbiology and Immunology, University of California, San Francisco 94143-0414, USA.
Blackburn
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EH
D000998:Antiviral Agents / Q000494:pharmacology; D001087:Arabinonucleosides / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004247:DNA / Q000235:genetics; D015224:Dideoxynucleosides / Q000494:pharmacology; D054306:Dideoxynucleotides; D004347:Drug Interactions; D017245:Foscarnet / Q000494:pharmacology; D006801:Humans; D015345:Oligonucleotide Probes; D012091:Repetitive Sequences, Nucleic Acid; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology*; D019098:Telomerase / Q000037:antagonists & inhibitors*; D016615:Telomere / Q000187:drug effects* / Q000235:genetics / Q000648:ultrastructure; D013942:Thymine Nucleotides / Q000494:pharmacology; D015215:Zidovudine / Q000031:analogs & derivatives / Q000494:pharmacology
D000998:Antiviral Agents; D001087:Arabinonucleosides; D015224:Dideoxynucleosides; D054306:Dideoxynucleotides; D015345:Oligonucleotide Probes; D018894:Reverse Transcriptase Inhibitors; D013942:Thymine Nucleotides; D017245:Foscarnet; C028771:9-arabinofuranosylguanine; D015215:Zidovudine; C052873:zidovudine triphosphate; C050220:2',3'-dideoxyguanosine; D004247:DNA; D019098:Telomerase