Identification of WTAP, a novel Wilms' tumour 1-associating protein
Identification of WTAP, a novel Wilms' tumour 1-associating protein
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DOI：
10.1093/oxfordjournals.hmg.a018914
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发表时间：
2000-09-22
影响因子：
3.5
通讯作者：
Davies, RC
中科院分区：
文献类型：
--
作者：
Little, NA;Hastie, ND;Davies, RC
Wilms肿瘤抑制基因WT1对泌尿生殖系统的正常发育至关重要，它似乎在某些细胞基因的转录和转录后调控中发挥作用。然而，尽管发现了许多潜在的靶基因，并分离了几种WT1结合蛋白，但WT1功能背后的机制尚不清楚。因此，本研究着手鉴定其他WT1相关蛋白，以帮助揭示WT1如何与细胞机制相互作用。我们报道了一种新的人类WT1相关蛋白WTAP，它是用酵母双杂交系统分离出来的。体外和体内实验表明，WTAP和WT1之间的相互作用是特异性的，并且是内源性的。从小鼠中分离出WTAP的同源基因，发现其在核苷酸和蛋白水平上的保守性为bb0 ~ 90%。利用荧光原位杂交技术将人类和小鼠的基因分别定位到6号染色体（被认为含有肿瘤抑制基因）和17号染色体的区域。研究表明，WTAP的表达模式是普遍存在的，可能反映了一种管家作用。WTAP是一种核蛋白，与WT1一样，它定位于整个核质和斑点中，并与剪接因子部分共定位。虽然这种相互作用的意义尚不清楚，但WTAP有望成为一个有趣的wt1结合伙伴。
The Wilms' tumour suppressor gene WT1 is essential for the normal development of the genitourinary system, It appears to play a role in both transcriptional and post-transcriptional regulation of certain cellular genes. However, the mechanisms behind WT1 function are not clearly understood despite the identification of numerous potential target genes and the isolation of several WT1-binding proteins. This study therefore sets out to identify other WT1-associating proteins to help to unravel how WT1 interacts with the cellular machinery. We report the identification of a novel human WT1-associating protein, WTAP, which was isolated using the yeast two-hybrid system, Both in vitro and in vivo assays have shown that the interaction between WTAP and WT1 is specific and occurs endogenously in cells. The mouse homologue of WTAP was isolated and found to be >90% conserved at the nucleotide and protein levels. The human and mouse genes were mapped using fluorescence in situ hybridization to regions in chromosomes 6 (which is thought to harbour a tumour suppressor gene) and 17, respectively. The expression pattern of WTAP was investigated and shown to be ubiquitous, perhaps reflecting a housekeeping role. WTAP is a nuclear protein, which like WT1 localizes throughout the nucleoplasm as well as in speckles and partially co-localizes with splicing factors. Although the significance of this interaction is not yet known, WTAP promises to be an interesting WT1-binding partner.