The HSV-1 ICP27 RGG box specifically binds flexible, GC-rich sequences but not G-quartet structures.
The HSV-1 ICP27 RGG box specifically binds flexible, GC-rich sequences but not G-quartet structures.
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DOI：
10.1093/nar/gkp793
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发表时间：
2009-11
影响因子：
14.9
通讯作者：
Sandri-Goldin RM
中科院分区：
文献类型：
--
作者：
Corbin-Lickfett KA;Chen IH;Cocco MJ;Sandri-Goldin RM
单纯疱疹病毒1 （HSV-1）蛋白ICP27是病毒基因表达的重要调控因子，通过n端RGG盒RNA结合基序直接识别和输出病毒RNA，这是RNA结合的必要和充分条件。表达包含RGG盒RNA结合基序的ICP27 n端肽，并利用EMSA和SELEX分析其结合特异性。在酵母三杂交分析中鉴定出与HSV-1糖蛋白C (gC) mRNA相对应的DNA寡核苷酸，在EMSA实验中筛选与ICP27 n端肽结合的DNA寡核苷酸。ICP27 n端能够结合大多数gC底物。值得注意的是，ICP27 RGG盒不能结合其他蛋白质的RGG结构域识别的g -四重奏结构。SELEX分析发现富含gc的RNA序列是识别的共同特征。SELEX和gC序列的NMR分析表明，能够与ICP27结合的序列没有形成二级结构，相反，不能与ICP27结合的序列给出了与碱基配对一致的光谱。因此，与其他RGG box蛋白相比，ICP27 RGG box在识别核酸序列方面具有独特性；它更喜欢柔性的、富含gc的基质，这些基质不会形成稳定的二级结构。
Herpes simplex virus 1 (HSV-1) protein ICP27, an important regulator for viral gene expression, directly recognizes and exports viral RNA through an N-terminal RGG box RNA binding motif, which is necessary and sufficient for RNA binding. An ICP27 N-terminal peptide, including the RGG box RNA binding motif, was expressed and its binding specificity was analyzed using EMSA and SELEX. DNA oligonucleotides corresponding to HSV-1 glycoprotein C (gC) mRNA, identified in a yeast three-hybrid analysis, were screened for binding to the ICP27 N-terminal peptide in EMSA experiments. The ICP27 N-terminus was able to bind most gC substrates. Notably, the ICP27 RGG box was unable to bind G-quartet structures recognized by the RGG domains of other proteins. SELEX analysis identified GC-rich RNA sequences as a common feature of recognition. NMR analysis of SELEX and gC sequences revealed that sequences able to bind to ICP27 did not form secondary structures and conversely, sequences that were not able to bind to ICP27 gave spectra consistent with base-pairing. Therefore, the ICP27 RGG box is unique in its recognition of nucleic acid sequences compared to other RGG box proteins; it prefers flexible, GC-rich substrates that do not form stable secondary structures.
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影响因子：
5.4
作者：
HARDY, WR;SANDRIGOLDIN, RM
通讯作者：
SANDRIGOLDIN, RM
影响因子：
10.5
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