Single-molecule FRET reveals the folding dynamics of the human telomerase RNA pseudoknot domain.
Single-molecule FRET reveals the folding dynamics of the human telomerase RNA pseudoknot domain.
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DOI：
10.1002/anie.201200526
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发表时间：
2012-06-11
影响因子：
16.6
通讯作者：
Stone, Michael D.
中科院分区：
文献类型：
--
作者：
Hengesbach, Martin;Kim, Nak-Kyoon;Feigon, Juli;Stone, Michael D.
端粒DNA的端粒酶催化合成为称为端粒的DNA-蛋白质结构提供了基础。端粒是逃避DNA加工所必需的，DNA加工可能导致线性染色体的溶核降解和融合。人端粒酶含有蛋白质逆转录酶（hTERT）、端粒酶RNA（hTR）和几种另外的蛋白质。[1]hTR在产生短端粒DNA重复序列的新型逆转录反应期间为hTERT提供模板序列。[2]此外，hTR为端粒酶相关蛋白的组装和功能提供了灵活的支架[3]，并且最近被认为有助于酶催化。[4]许多致病性突变已被定位到hTR上;[5]然而这些突变的确切机制仍不清楚。在体外将hTR与hTERT重组为催化活性酶需要两个可分离的RNA结构域，其中包含几个保守的结构基序，包括RNA假结（PK）折叠。[5-10]野生型和突变型最小PK构建体的NMR研究揭示了折叠的假k-not中的三级相互作用，其形成保守的三螺旋以及部分折叠的发夹。[10-15]单分子实验已经使用机械力来探测最小PK构建体的折叠/解折叠[11]，以及改良的双色重合检测测定（TCCD-lex）来重建溶液中较大hTR PK结构域构建体的RNA构象分布。[16]由于天然hTR PK结构域的大尺寸，其折叠特性的分析迄今为止在很大程度上限于天然RNA序列的截短变体。为了规避这个
Telomerase catalyzed synthesis of telomere DNA provides the foundation for DNA-protein structures called telomeres. Telomeres are required to evade DNA processing which may result in nucleolytic degradation and fusion of linear chromosomes. Human telomerase contains a protein reverse transcriptase (hTERT), telomerase RNA (hTR), and several additional proteins.[1] hTR provides the template sequence for hTERT during a novel reverse transcription reaction that produces short telomere DNA repeat sequences.[2] Additionally, hTR provides a flexible scaffold for the assembly and function of telomerase associated proteins [3] and has recently been suggested to contribute to enzyme catalysis.[4] Many pathogenic mutations have been mapped to hTR;[5] however the precise mechanisms of these mutations remain unclear.In vitro reconstitution of hTR with hTERT into a catalytically active enzyme requires two separable RNA domains harboring several conserved structural motifs, including an RNA pseudoknot (PK) fold.[5–10] NMR studies of wild-type and mutant minimal PK constructs have revealed tertiary interactions in the folded pseudok-not that form a conserved triple helix, as well as a partially folded hairpin.[10–15] Single molecule experiments have used mechanical force to probe the folding/unfolding of a minimal PK construct [11], and a modified two color coincidence detection assay (TCCD-lex) to reconstruct the distribution of RNA conformations for a larger hTR PK domain construct in solution.[16] Due to the large size of the native hTR PK domain, analyses of its folding properties have thus far been largely limited to truncated variants of the natural RNA sequence. To circumvent this
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影响因子：
5.3
作者：
Egan, Emily D.;Collins, Kathleen
通讯作者：
Collins, Kathleen
影响因子：
16
作者：
Theimer, CA;Blois, CA;Feigon, J
通讯作者：
Feigon, J
影响因子：
5.6
作者：
Kim, Nak-Kyoon;Zhang, Qa;Feigon, Juli
通讯作者：
Feigon, Juli
影响因子：
9.2
作者：
Beattie, TL;Zhou, W;Harrington, L
通讯作者：
Harrington, L
DOI：
10.1073/pnas.262663599
发表时间：
2002-12-24
影响因子：
11.1
作者：
Comolli, LR;Smirnov, I;James, TL
通讯作者：
James, TL