Soft Interactions with Model Crowders and Non-canonical Interactions with Cellular Proteins Stabilize RNA Folding.
Soft Interactions with Model Crowders and Non-canonical Interactions with Cellular Proteins Stabilize RNA Folding.
复制标题
DOI：
10.1016/j.jmb.2017.10.030
复制
发表时间：
2018-02-16
影响因子：
5.6
通讯作者：
Walter NG
中科院分区：
文献类型：
--
作者：
Daher M;Widom JR;Tay W;Walter NG
活细胞含有不同的生物聚合物，创造了一个异质拥挤的环境，其对RNA折叠的影响知之甚少。在这里，我们已经使用单分子荧光共振能量转移来监测发夹状核酶的三级结构形成，作为一个模型来探测聚乙二醇（PEG）和酵母细胞提取物作为拥挤剂的影响。正如预期的那样，PEG稳定的对接，催化活性状态的核酶，部分通过排除体积效应;出乎意料的是，我们发现的证据表明，它还显示软，非特异性的相互作用与核酶。酵母提取物在蛋白质浓度比细胞内低1,000倍时对折叠有深远的影响，表明特异性相互作用超过体积排阻。凝胶迁移试验和亲和下拉质谱法确定了许多非典型的RNA结合蛋白，稳定核酶折叠，这些无处不在的蛋白质的表观伴侣活性显着补偿低counterweight环境的细胞。
Living cells contain diverse biopolymers, creating a heterogeneous crowding environment, the impact of which on RNA folding is poorly understood. Here, we have used single-molecule fluorescence resonance energy transfer to monitor tertiary structure formation of the hairpin ribozyme as a model to probe the effects of polyethylene glycol (PEG) and yeast cell extract as crowding agents. As expected, PEG stabilizes the docked, catalytically active state of the ribo-zyme, in part through excluded volume effects; unexpectedly, we found evidence that it additionally displays soft, non-specific interactions with the ribozyme. Yeast extract has a profound effect on folding at protein concentrations 1,000-fold lower than found intracellularly, suggesting the dominance of specific interactions over volume exclusion. Gel shift assays and affinity pull-down followed by mass spectrometry identified numerous non-canonical RNA-binding proteins that stabilize ribozyme folding; the apparent chaperoning activity of these ubiquitous proteins significantly compensates for the low-counterion environment of the cell.
登录
查看更多内容
DOI：
10.1016/j.tem.2015.09.012
发表时间：
2015-12
期刊：
Trends in endocrinology and metabolism: TEM
影响因子：
--
作者：
Castello A;Hentze MW;Preiss T
通讯作者：
Preiss T
影响因子：
16.6
作者：
Beckmann BM;Horos R;Fischer B;Castello A;Eichelbaum K;Alleaume AM;Schwarzl T;Curk T;Foehr S;Huber W;Krijgsveld J;Hentze MW
通讯作者：
Hentze MW
弥合体外和体内RNA折叠之间的间隙。
影响因子：
6.1
作者：
Leamy KA;Assmann SM;Mathews DH;Bevilacqua PC
通讯作者：
Bevilacqua PC
影响因子：
4.4
作者：
LEBOWITZ, JL;HELFAND, E;PRAESTGAARD, E
通讯作者：
PRAESTGAARD, E
影响因子：
15
作者：
Kilburn, Duncan;Roh, Joon Ho;Guo, Liang;Briber, Robert M.;Woodson, Sarah A.
通讯作者：
Woodson, Sarah A.