Targeted 2'-O methylation at a nucleotide within the pseudoknot of telomerase RNA reduces telomerase activity in vivo.

Molecular and Cellular Biology
Chao Huang, Yi-Tao Yu

Abstract

Telomerase RNA is an essential component of telomerase, a ribonucleoprotein enzyme that maintains chromosome ends in most eukaryotes. Here we employ a novel approach, namely, RNA-guided RNA modification, to assess whether introducing 2'-O methylation into telomerase RNA can influence telomerase activity in vivo. We generate specific 2'-O methylation sites in and adjacent to the triple helix (within the conserved pseudoknot structure) of Saccharomyces cerevisiae telomerase RNA (TLC1). We show that 2'-O methylation at U809 reduces telomerase activity, resulting in telomere shortening, whereas 2'-O methylation at A804 or A805 leads to moderate telomere lengthening. Importantly, we also show that targeted 2'-O methylation does not affect TLC1 levels and that 2'-O-methylated TLC1 appears to be efficiently assembled into telomerase ribonucleoprotein. Our results demonstrate that RNA-guided RNA modification is a highly useful approach for modulating telomerase activity.

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Citations

Mar 31, 2012·Nature Protocols·Chao HuangYi-Tao Yu
Nov 30, 2011·Human Gene Therapy·Anne-Marie W TurnerKevin V Morris
Feb 9, 2016·Methods : a Companion to Methods in Enzymology·Chao HuangYi-Tao Yu
Oct 12, 2013·Wiley Interdisciplinary Reviews. RNA·Nicholas K Conrad
Apr 16, 2015·PloS One·Isioma I EnweremMichael D Hebert
Apr 3, 2013·Current Protocols in Molecular Biology·Chao Huang, Yi-Tao Yu

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