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A bulged stem tethers Est1p to telomerase RNA in budding yeast
A bulged stem tethers Est1p to telomerase RNA in budding yeast.
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It is well established that the template for telomeric DNA synthesis is provided by the RNA subunit of telomerase; however, the additional functions provided by most of the rest of the RNA (>1000 nucleotides in budding yeast) are largely unknown. By alignment of telomerase RNAs of Saccharomyces cerevisiae and six Kluyveromyces species followed by mutagenesis of the S. cerevisiae RNA, we found a conserved region that is essential for telomere maintenance. Phylogenetic analysis and computer folding revealed that this region is conserved not only in primary nucleotide sequence but also in secondary structure. A common bulged-stem structure was predicted in all seven yeast species. Mutational analysis showed the structure to be essential for telomerase function. Suppression of bulged-stem mutant phenotypes by overexpression of Est1p and loss of co-immunoprecipitation of the mutant RNAs with Est1p indicated that this bulged stem is necessary for association of Est1p, a telomerase regulatory subunit. Est1p in yeast extract bound specifically to a small RNA containing the bulged stem, suggesting a direct interaction. We propose that this RNA structure links the enzymatic core of telomerase with Est1p, thereby allowing Est1p to recruit or activate telomerase at the telomere.
Seto AG, Livengood AJ, Tzfati Y, Blackburn EH, Cech TR
Genes & development
2002-11-01 00:00
16
21
2800-12
Conserved Sequence,Mutation,RNA, Fungal,Saccharomyces cerevisiae,Saccharomyces cerevisiae Proteins,Sequence Alignment,Sequence Analysis,Telomerase,EST1 protein, S cerevisiae,RNA, Fungal,Saccharomyces cerevisiae Proteins,Telomerase
Department of Chemistry and Biochemistry, University of Colorado, Boulder, Colorado 80309-0215, USA
Genes Dev.
NIGMS GM26259, NIGMS GM28039
0890-9369
10.1101/gad.1029302
125
True
12414733
