Rapid protein sequencing by tandem mass spectrometry and cDNA cloning of p20-CGGBP - A novel protein that binds to the unstable triplet repeat 5'-d(CGG)(n)-3' in the human FMR1 gene
Rapid protein sequencing by tandem mass spectrometry and cDNA cloning of p20-CGGBP - A novel protein that binds to the unstable triplet repeat 5'-d(CGG)(n)-3' in the human FMR1 gene
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DOI：
10.1074/jbc.272.27.16761
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发表时间：
1997-07-04
影响因子：
4.8
通讯作者：
Doerfler, W
中科院分区：
文献类型：
--
作者：
Deissler, H;Wilm, M;Doerfler, W
人类FMR1基因5'-未翻译区不稳定的5‘-d(CGG)(n)-3’重复序列的自主扩增导致脆性X综合征，这是人类男性智力迟钝的最常见原因之一。我们最近描述了一种蛋白质p20-CGGBP的分离，该蛋白质特异性地结合双链三核苷酸重复序列5'-d(CGG)-3' (Deissler， H., Behn-Krappa, a ., and Doerfler， W. (1996) J. Biol。我们现在证明p20-CGGBP也可以与中断的重复序列结合，通过纳米电喷雾质谱法获得p20-CGGBP的肽序列标签，对表达的序列标签数据库进行筛选，检索包含p20-CGGBP全长编码序列的克隆。细菌表达的融合蛋白p20-CGGBP- 6xhis显示出与真实的p20-CGGBP相似的双链5‘-d(CGG),-3’重复的结合模式，该新蛋白与其他已知蛋白缺乏整体同源性，但携带假定的核定位信号，p20-CGGBP基因在哺乳动物中保守，但与非脊椎动物物种无同源性，编码新蛋白序列的基因已定位到人类3号染色体。
The autonomous expansion of the unstable 5'-d(CGG)(n)-3' repeat in the 5'-untranslated region of the human FMR1 gene leads to the fragile X syndrome, one of the most frequent causes of mental retardation in human males, We have recently described the isolation of a protein p20-CGGBP that binds sequence-specifically to the double stranded trinucleotide repeat 5'-d(CGG)-3' (Deissler, H., Behn-Krappa, A., and Doerfler, W. (1996) J. Biol. Chem. 271, 4327-4334), We demonstrate now that the p20-CGGBP can also bind to an interrupted repeat sequence, Peptide sequence tags of p20-CGGBP obtained by nanoelectrospray mass spectrometry were screened against an expressed sequence tag data base, retrieving a clone that contained the full-length coding sequence for p20-CGGBP. A bacterially expressed fusion protein p20-CGGBP-6xHis exhibits a binding pattern to the double-stranded 5'-d(CGG),-3' repeat similar to that of the authentic p20-CGGBP, This novel protein lacks any overall homology to other known proteins but carries a putative nuclear localization signal, The p20-CGGBP gene is conserved among mammals but shows no homology to non-vertebrate species, The gene encoding the sequence for the new protein has been mapped to human chromosome 3.