Analysis of the structure of human telomerase RNA in vivo.
Telomerase is a ribonucleoprotein reverse transcriptase that synthesises telomeric DNA. The RNA component of telomerase acts as a template for telomere synthesis and binds the reverse transcriptase. In this study, we have performed in vivo and in vitro structural analyses of human telomerase RNA (hTR). In vivo mapping experiments showed that the 5'-terminal template domain of hTR folds into a long hairpin structure, in which the template sequence occupies a readily accessible position. Intriguingly, neither in vivo nor in vitro mapping of hTR confirmed formation of a stable 'pseudoknot' helix, suggesting that this functionally essential long range interaction is formed only temporarily. In vitro control mappings demonstrated that the 5'-terminal template domain of hTR cannot fold correctly in the absence of cellular protein factors. The 3'-terminal domain of hTR, both in vivo and in vitro, folds into the previously predicted box H/ACA snoRNA-like 'hairpin-hinge-hairpin-tail' structure. Finally, comparison of the in vivo and in vitro modification patterns of hTR revealed several regions that might be directly involved in binding of telomerase reverse transcriptase or other telomerase proteins.
Antal
Mária
M
Biological Research Center, Hungarian Academy of Sciences, PO Box 521, H-6701 Szeged, Hungary.
Boros
Eva
E
Solymosy
Ferenc
F
Kiss
Tamás
T
D017124:Conserved Sequence; D004720:Endonucleases / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008957:Models, Genetic; D009690:Nucleic Acid Conformation; D011560:Pseudouridine / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D020537:RNA, Small Nucleolar / Q000737:chemistry; D019098:Telomerase / Q000737:chemistry*; D013698:Templates, Genetic
D020537:RNA, Small Nucleolar; C422952:telomerase RNA; D011560:Pseudouridine; D012313:RNA; D019098:Telomerase; D004720:Endonucleases