Telomerase limits the extent of base pairing between template RNA and telomeric DNA

被引:52
作者
Förstemann, K
Lingner, J [1 ]
机构
[1] Ecole Polytech Fed Lausanne, Swiss Inst Expt Canc Res, CH-1066 Epalinges, Switzerland
[2] Natl Ctr Competence Res Frontiers Genet, CH-1066 Epalinges, Switzerland
关键词
telomerase reaction cycle; telomerase RNA structure; dimethyl sulphate; reverse transcriptase;
D O I
10.1038/sj.embor.7400374
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Telomerase is the ribonucleoprotein reverse transcriptase that adds telomeric DNA repeats to the ends of chromosomes. This involves annealing of the telomerase RNA template to the 30 end of the chromosome, reverse transcription of the RNA template by the telomerase reverse transcriptase polypeptide and translocation. Here, we overexpress and partially purify the catalytically active yeast telomerase core in its natural host and probe telomerase RNA base methylation accessibility with dimethyl sulphate in the presence and absence of a DNA substrate and after substrate elongation. The length of the RNA - DNA hybrid is kept constant at seven base pairs after primer binding and elongation. Thus, new base-pair formation at the 30 end of the substrate during elongation coincides with disruption of base-pair interactions at the other side of the template. Presumably, this circumvents the generation of an exceedingly high energy barrier for translocation and dissociation. Our analysis also corroborates recently proposed yeast telomerase RNA secondary structure models.
引用
收藏
页码:361 / 366
页数:6
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