The fragile X mental retardation protein interacts with U-rich RNAs in a yeast three-hybrid system
The fragile X mental retardation protein interacts with U-rich RNAs in a yeast three-hybrid system
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DOI：
10.1016/s0006-291x(03)00766-6
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发表时间：
2003-05-30
影响因子：
3.1
通讯作者：
Denman, RB
中科院分区：
文献类型：
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作者：
Dolzhanskaya, N;Sung, YJ;Denman, RB
我们最近发现了几个EST，结合脆性X智力低下蛋白（FMRP）在体外。为了确定它们是否在体内相互作用，我们在酿酒酵母组氨酸营养缺陷型中进行了三杂交筛选。我们证明，两个EST支持生长的组氨酸和β-半乳糖苷酶的活性时，共表达FMRP在选择性生长条件下。相反，铁反应元件（IRE）RNA则没有。同样，当与铁反应元件结合蛋白（IRP）共表达时，EST不支持生长或β-半乳糖苷酶活性。每个EST相对较小，与FMR 1 mRNA中含有FMRP结合决定簇的序列具有40%的同一性。有趣的是，虽然EST都不包含G-四联体结构基序，但它们确实包含在mRNA中发现的富含U的序列，这些序列在体外结合和体内与FMRP结合。这表明富U元素包含FMRP识别的另一个基序。(C)2003 Elsevier Science（美国）。All rights reserved.
We recently identified several ESTs that bind to the fragile X mental retardation protein (FMRP) in vitro. To determine whether they interacted in vivo we performed three-hybrid screens in a Saccharomyces cerevisiae histidine auxotroph. We demonstrate that two of the ESTs support growth on histidine and transduce beta-galactosidase activity when co-expressed with FMRP under selective growth conditions. In contrast, the iron response element (IRE) RNA does not. Likewise, the ESTs do not support growth or transduce beta-galactosidase activity when co-expressed with the iron response element binding protein (IRP). Each EST is relatively small and has 40% identity with a sequence in FMR1 mRNA harboring FMRP binding determinants. Interestingly, while neither the ESTs contain a G-quartet structural motif they do contain U-rich sequences that are found in mRNA with demonstrated in vitro binding and in vivo association with FMRP. This indicates that U-rich elements comprise another motif recognized by FMRP. (C) 2003 Elsevier Science (USA). All rights reserved.