Structural and functional implications of the QUA2 domain on RNA recognition by GLD-1.
Structural and functional implications of the QUA2 domain on RNA recognition by GLD-1.
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DOI：
10.1093/nar/gku445
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发表时间：
2014-07
影响因子：
14.9
通讯作者：
Allain FH
中科院分区：
文献类型：
--
作者：
Daubner GM;Brümmer A;Tocchini C;Gerhardy S;Ciosk R;Zavolan M;Allain FH
星星家族包括在RNA调节过程中起关键作用的核糖核酸（RNA）结合蛋白。RNA识别是通过一个KH结构域与一个额外的α-螺旋（QUA 2）来实现的，该结构域似乎将SF 1（智人）的RNA结合表面扩展到6个核苷酸，并将GLD-1（秀丽隐杆线虫）的RNA结合表面扩展到7个核苷酸。为了理解这种可能的特异性差异的结构基础，我们用tra-2基因中鉴定的完整共有序列确定了GLD-1 KH-QUA 2的溶液结构。与SF 1相比，GLD-1 KH-QUA 2界面采用了不同的构象，从而在5′端确实产生了额外的尿嘧啶序列特异性结合口袋。我们的生物信息学分析强调了该结合口袋的功能相关性，表明具有该5′端尿嘧啶的GLD-1结合位点更能预测信使RNA对gld-1敲除的功能反应。我们进一步揭示了体外KH-QUA 2界面的重要性，并且其在体内的改变影响了依赖于GLD-1结合基序序列的翻译抑制水平。总之，我们证明了QUA 2结构域将GLD-1与星星家族的其他成员区分开来，并且更普遍地有助于调节含有KH结构域的蛋白质的RNA结合亲和力和特异性。
The STAR family comprises ribonucleic acid (RNA)-binding proteins that play key roles in RNA-regulatory processes. RNA recognition is achieved by a KH domain with an additional α-helix (QUA2) that seems to extend the RNA-binding surface to six nucleotides for SF1 (Homo sapiens) and seven nucleotides for GLD-1 (Caenorhabditis elegans). To understand the structural basis of this probable difference in specificity, we determined the solution structure of GLD-1 KH-QUA2 with the complete consensus sequence identified in the tra-2 gene. Compared to SF1, the GLD-1 KH-QUA2 interface adopts a different conformation resulting indeed in an additional sequence-specific binding pocket for a uracil at the 5′end. The functional relevance of this binding pocket is emphasized by our bioinformatics analysis showing that GLD-1 binding sites with this 5′end uracil are more predictive for the functional response of the messenger RNAs to gld-1 knockout. We further reveal the importance of the KH-QUA2 interface in vitro and that its alteration in vivo affects the level of translational repression dependent on the sequence of the GLD-1 binding motif. In conclusion, we demonstrate that the QUA2 domain distinguishes GLD-1 from other members of the STAR family and contributes more generally to the modulation of RNA-binding affinity and specificity of KH domain containing proteins.
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影响因子：
2.7
作者：
Cordier, F;Dingley, AJ;Grzesiek, S
通讯作者：
Grzesiek, S
影响因子：
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作者：
Daubner, Gerrit M.;Clery, Antoine;Allain, Frederic H-T
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Allain, Frederic H-T
影响因子：
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作者：
Beuck, Christine;Qu, Song;Fagg, W. Samuel;Ares, Manuel, Jr.;Williamson, James R.
通讯作者：
Williamson, James R.
影响因子：
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作者：
Carmel AB;Wu J;Lehmann-Blount KA;Williamson JR
通讯作者：
Williamson JR
影响因子：
5.6
作者：
Herrmann, T;Güntert, P;Wüthrich, K
通讯作者：
Wüthrich, K