The tetranucleotide UCAY directs the specific recognition of RNA by the Nova K-homology 3 domain
The tetranucleotide UCAY directs the specific recognition of RNA by the Nova K-homology 3 domain
复制标题
DOI：
10.1073/pnas.090553997
复制
发表时间：
2000-05-23
影响因子：
11.1
通讯作者：
Darnell, RB
中科院分区：
文献类型：
--
作者：
Jensen, KB;Musunuru, K;Darnell, RB
Nova家族蛋白是自身免疫性疾病副肿瘤性眼阵挛-肌阵挛共济失调的靶抗原，含有K-同源(KH)型RNA结合域。最近，Nova-1蛋白被证明通过与包含四核苷酸UCAU重复的内含子元件结合来调节α2甘氨酸受体亚单位前mRNA的选择性剪接。在这里，我们使用选择扩增来证明Nova的Kh3结构域在20碱基的发夹RNA的背景下识别单个UCAY元件；UCAY四核苷酸最佳地作为发夹支架的环元件存在，并且需要蛋白质残基的C-末端到先前定义的KH结构域。这些结果表明，KH结构域一般识别四核苷酸基序，KH结构域的生物RNA靶标可能使用RNA二级结构或重复序列元件来实现蛋白质结合的高亲和力和特异性。
The Nova family of proteins are target antigens in the autoimmune disorder paraneoplastic opsoclonus-myoclonus ataxia and contain K-homology (KH)-type RNA binding domains. The Nova-1 protein has recently been shown to regulate alternative splicing of the alpha 2 glycine receptor subunit pre-mRNA by binding to an intronic element containing repeats of the tetranucleotide UCAU, Here, we have used selection-amplification to demonstrate that the KH3 domain of Nova recognizes a single UCAY element in the context of a 20-base hairpin RNA; the UCAY tetranucleotide is optimally presented as a loop element of the hairpin scaffold and requires protein residues C-terminal to the previously defined KH domain. These results suggest that KH domains in general recognize tetranucleotide motifs and that biological RNA targets of KH domains may use either RNA secondary structure or repeated sequence elements to achieve high affinity and specificity of protein binding.