Sequence determinants for the tandem recognition of UGU and CUG rich RNA elements by the two N--terminal RRMs of CELF1.
Sequence determinants for the tandem recognition of UGU and CUG rich RNA elements by the two N--terminal RRMs of CELF1.
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DOI：
10.1093/nar/gkr510
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发表时间：
2011-10
影响因子：
14.9
通讯作者：
Emsley J
中科院分区：
文献类型：
--
作者：
Edwards J;Malaurie E;Kondrashov A;Long J;de Moor CH;Searle MS;Emsley J
CUGBP，Elav-like family member 1（CELF 1）是一种RNA结合蛋白，在剪接、mRNA降解和翻译的调节中具有重要作用。CELF 1含有三个RNA识别基序（RRM）。我们使用凝胶阻滞，凝胶过滤，等温滴定量热法和NMR滴定研究，以调查的前两个RRM CELF 1的RNA的识别。NMR显示RRM 1在结合UGU和CUG重复序列时是混杂的，具有可比的化学位移扰动。相比之下，RRM 2对UGUU而不是CUG基序显示出更大的选择性。系统地研究了含有两个结合结构域（RRM 1和RRM 2）的构建体（T187）与具有不同长度接头序列UGU（U）xUGU（其中x = 1-7）的串联UGU RNA结合位点的相互作用。单个U间隔区仅与RRM 1相互作用，表明在相邻位点同时容纳两个RRM的空间限制，以及RRM之间结合亲和力的细微差异。然而，对于x = 2-4的RNA序列，高亲和力协同结合（Kd ~ 0.4 μM）是明显的，但较长的间隔区（x ≥ 5）导致亲和力降低10倍。我们的分析合理化的高亲和力相互作用的T187与11聚体的GRE共识调控序列UGUUUGUUGU和CELF 1结合位点的预测有显着的后果。
CUGBP, Elav-like family member 1 (CELF1) is an RNA binding protein with important roles in the regulation of splicing, mRNA decay and translation. CELF1 contains three RNA recognition motifs (RRMs). We used gel retardation, gel filtration, isothermal titration calorimetry and NMR titration studies to investigate the recognition of RNA by the first two RRMs of CELF1. NMR shows that RRM1 is promiscuous in binding to both UGU and CUG repeat sequences with comparable chemical shift perturbations. In contrast, RRM2 shows greater selectivity for UGUU rather than CUG motifs. A construct (T187) containing both binding domains (RRM1 and RRM2) was systematically studied for interaction with tandem UGU RNA binding sites with different length linker sequences UGU(U)xUGU where x = 1–7. A single U spacer results in interactions only with RRM1, demonstrating both steric constraints in accommodating both RRMs simultaneously at adjacent sites, and also subtle differences in binding affinities between RRMs. However, high affinity co-operative binding (Kd ~ 0.4 µM) is evident for RNA sequences with x = 2–4, but longer spacers (x ≥ 5) lead to a 10-fold reduction in affinity. Our analysis rationalizes the high affinity interaction of T187 with the 11mer GRE consensus regulatory sequence UGUUUGUUUGU and has significant consequences for the prediction of CELF1 binding sites.
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影响因子：
5.3
作者：
Baldwin, BR;Timchenko, NA;Zahnow, CA
通讯作者：
Zahnow, CA
DOI：
10.1016/j.bbrc.2006.06.177
发表时间：
2006-09-01
影响因子：
3.1
作者：
Barreau, Carine;Watrin, Tanguy;Paillard, Luc
通讯作者：
Paillard, Luc
影响因子：
11.4
作者：
de Moor, CH;Richter, JD
通讯作者：
Richter, JD
影响因子：
5.3
作者：
Kress, Chantal;Gautier-Courteille, Carole;Paillard, Luc
通讯作者：
Paillard, Luc
影响因子：
14.9
作者：
Delaunay, J;Mée, GL;Aït-Ahmed, O
通讯作者：
Aït-Ahmed, O