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首页> 外文期刊>Journal of Molecular Biology >Telomerase activity is sensitive to subtle perturbations of the TLC1 pseudoknot 3′ stem and tertiary structure
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Telomerase activity is sensitive to subtle perturbations of the TLC1 pseudoknot 3′ stem and tertiary structure

机译：端粒酶活性对TLC1假结3'茎和三级结构的微扰很敏感

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摘要

Pseudoknot formation in the core region of the telomerase RNA has been demonstrated to be important for telomerase activity in vertebrates, ciliates, and yeast. Characterization of the Saccharomyces cerevisiae telomerase RNA (TLC1) pseudoknot identified tertiary structural interactions that are also important for telomerase activity, as previously observed for the Kluyveromyces lactis and human telomerase RNA pseudoknots. In addition, the contributions of backbone ribose 2′-OH groups in the pseudoknot to telomerase catalysis were investigated previously, using 2′-OH (ribose) to 2′-H (deoxyribose) or 2′-O-methyl substitutions in the stem 2 helix, and it was proposed that one or more 2′-OH groups from the stem 2 sequences at or near the triple helix participate in telomerase catalysis. Based on these studies and investigations of the structural and thermodynamic properties of the TLC1 RNA pseudoknot region, we have examined the structural and thermodynamic perturbations of the 2′-O-methyl and 2′-H substituted pseudoknots, using UV-monitored thermal denaturation, native gel electrophoresis, and circular dichroism spectroscopy. Our results demonstrate the presence of A-form helical geometry perturbations in the backbone sugar substituted pseudoknots, show a correlation between thermodynamic stability and telomerase activity, and are consistent with the identification of the U809 ribose 2′-OH as a potential contributor to telomerase activity.
机译：已经证明在端粒酶RNA的核心区域中的假结形成对于脊椎动物，纤毛虫和酵母中的端粒酶活性是重要的。酿酒酵母端粒酶RNA（TLC1）假结的特征鉴定了三级结构相互作用，这对于端粒酶活性也很重要，如先前对乳酸克鲁维酵母和人端粒酶RNA假结所观察到的。此外，先前使用茎中的2'-OH（核糖）至2'-H（脱氧核糖）或2'-O-甲基取代，研究了假结中的主链核糖2'-OH基对端粒酶催化的贡献。 2个螺旋，并且提出了来自三螺旋或附近的茎2序列的一个或多个2'-OH基团参与端粒酶催化。基于这些研究和对TLC1 RNA假结区域的结构和热力学性质的研究，我们使用紫外线监测的热变性研究了2'-O-甲基和2'-H取代的假结的结构和热力学扰动，天然凝胶电泳和圆二色谱。我们的结果表明，主链糖取代的假结中存在A型螺旋几何扰动，表明热力学稳定性与端粒酶活性之间存在相关性，并且与U809核糖2'-OH鉴定为端粒酶活性的潜在贡献者是一致的。

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