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| Item Type: | Article |
|---|---|
| Title: | Telomerase protein rather than its RNA is the target of phosphorothioate-modified oligonucleotides |
| Creators Name: | Matthes, E. and Lehmann, C. |
| Abstract: | Human telomerase is a ribonucleoprotein which uses its internal RNA moiety as a template for telomeric DNA synthesis. This enzyme is up-regulated in most malignant tumors and is therefore considered as a possible cancer target. Here we examined the effects of differently modified oligomers on telomeraseactivity from HL-60 cell extracts (TRAP-ezetrade mark assay). Phosphorothioate-modified oligonucleotides (PS-ODNs) inhibited telomerase activity at subnanomolar concen-trations and proved to be more efficient than peptide nucleic acids. In contrast to all the investigated oligomers, PS-ODNs were found to bind to the protein motif of telomerase called the primer binding site but poorly to its RNA. This is suggested by kinetic investigations demonstrating a competitive interaction of PS-ODNs and TS primer at the primer binding site. The K m value of the TS primer was 10.8 nM, the K i value of a 20mer PS-ODN was 1.6 nM. When the TS primer was PS-modified a striking increase in the telomerase activity was found which correlates with the number of phosphodiesters replaced. The K m value of a completely PS-modified TS primer was 0.56 nM. Based on these results the design of chimeric ODNs is proposed consisting of a 5'-PS-modified part targeting the primer binding site and a 3'-terminus part targeting the telomerase RNA. |
| Keywords: | Antisense Oligodeoxyribonucleotides, Competitive Binding, Cultured Tumor Cells, DNA Primers, HL-60 Cells, RNA, Structure-Activity Relationship, Telomerase, Thionucleotides |
| Source: | Nucleic Acids Research |
| ISSN: | 0305-1048 |
| Publisher: | Oxford University Press |
| Volume: | 27 |
| Number: | 4 |
| Page Range: | 1152-1158 |
| Date: | 15 February 1999 |
| Official Publication: | http://nar.oxfordjournals.org/cgi/content/abstract/27/4/1152 |
| PubMed: | View item in PubMed |
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