The Runt domain of AML1 (RUNX1) binds a sequence-conserved RNA motif that mimics a DNA element.
The Runt domain of AML1 (RUNX1) binds a sequence-conserved RNA motif that mimics a DNA element.
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DOI：
10.1261/rna.037879.112
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发表时间：
2013-07
期刊：
影响因子：
--
通讯作者：
Kozu T
中科院分区：
文献类型：
--
作者：
Fukunaga J;Nomura Y;Tanaka Y;Amano R;Tanaka T;Nakamura Y;Kawai G;Sakamoto T;Kozu T
AML 1（RUNX 1）是造血的关键转录因子。获得特异性结合AML 1 Runt结构域的RNA适体。所有分离的适体均含有保守的序列基序5′-NNCCAC-3′和5′-GCGMGN′N′-3′（M：A或C; N和N′形成Watson-Crick碱基对）。基序的三个鸟嘌呤直接被Runt结构域的三个精氨酸残基识别。AML 1（RUNX 1）是造血的关键转录因子，通过其N-末端Runt结构域与靶基因的Runt结合双链DNA元件（RDE）结合。AML 1基因的异常在人类白血病中经常被发现。为了更好地了解AML 1及其在诊断和治疗中的潜在用途，我们获得了特异性结合AML 1 Runt结构域的RNA适体。酶探测和核磁共振分析显示，Apt 1-S是其中一种适体的截短变体，具有CACG四环和两个由内环隔开的茎区。所有分离的核酸适体均含有5′-NNCCAC-3′和5′-GCGMGN′N′-3′（M：A或C; N和N′形成Watson-Crick碱基对）保守序列基序。基序包含一个AC错配和一个碱基凸出。Apt 1-S的突变分析表明，基序的三个鸟嘌呤是重要的Runt结合是RDE的三个鸟嘌呤，这是由Runt结构域的三个精氨酸残基直接识别。Runt结构域的突变分析显示，用于Apt 1-S结合的氨基酸残基与用于RDE结合的氨基酸残基相似。此外，适体在体外与RDE竞争结合Runt结构域。这些结果表明，AML 1蛋白的Runt结构域与模拟DNA的适体的基序结合。我们的研究结果将为RNA在基础科学和应用科学中的功能和效用提供新的见解。
AML1 (RUNX1) is a key transcription factor for hematopoiesis. RNA aptamers that bind specifically to the AML1 Runt domain were obtained. All the isolated aptamers contain the conserved sequence motif 5′-NNCCAC-3′ and 5′-GCGMGN′N′-3′ (M:A or C; N and N′ form Watson–Crick base pairs). Three guanines of the motif are directly recognized by three arginine residues of the Runt domain. AML1 (RUNX1) is a key transcription factor for hematopoiesis that binds to the Runt-binding double-stranded DNA element (RDE) of target genes through its N-terminal Runt domain. Aberrations in the AML1 gene are frequently found in human leukemia. To better understand AML1 and its potential utility for diagnosis and therapy, we obtained RNA aptamers that bind specifically to the AML1 Runt domain. Enzymatic probing and NMR analyses revealed that Apt1-S, which is a truncated variant of one of the aptamers, has a CACG tetraloop and two stem regions separated by an internal loop. All the isolated aptamers were found to contain the conserved sequence motif 5′-NNCCAC-3′ and 5′-GCGMGN′N′-3′ (M:A or C; N and N′ form Watson–Crick base pairs). The motif contains one AC mismatch and one base bulged out. Mutational analysis of Apt1-S showed that three guanines of the motif are important for Runt binding as are the three guanines of RDE, which are directly recognized by three arginine residues of the Runt domain. Mutational analyses of the Runt domain revealed that the amino acid residues used for Apt1-S binding were similar to those used for RDE binding. Furthermore, the aptamer competed with RDE for binding to the Runt domain in vitro. These results demonstrated that the Runt domain of the AML1 protein binds to the motif of the aptamer that mimics DNA. Our findings should provide new insights into RNA function and utility in both basic and applied sciences.
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影响因子：
2.9
作者：
FAVOROVA, OO;FASIOLO, F;EBEL, JP
通讯作者：
EBEL, JP
影响因子：
5.4
作者：
MELNIKOVA, IN;CRUTE, BE;SPECK, NA
通讯作者：
SPECK, NA
影响因子：
64.8
作者：
Davis, Brandi N.;Hilyard, Aaron C.;Hata, Akiko
通讯作者：
Hata, Akiko
DOI：
10.1038/86264
发表时间：
2001-04-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Bravo, J;Li, Z;Warren, AJ
通讯作者：
Warren, AJ
影响因子：
2.9
作者：
Endo, Kei;Nakamura, Yoshikazu
通讯作者：
Nakamura, Yoshikazu