Molecular analysis of a novel winged helix protein, WIN - Expression pattern, DNA binding property, and alternative splicing within the DNA binding domain
Molecular analysis of a novel winged helix protein, WIN - Expression pattern, DNA binding property, and alternative splicing within the DNA binding domain
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DOI：
10.1074/jbc.272.32.19827
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发表时间：
1997-08-08
影响因子：
4.8
通讯作者：
Wong, GG
中科院分区：
文献类型：
--
作者：
Yao, KM;Sha, M;Wong, GG
我们从大鼠胰岛素瘤细胞系INS-1中克隆了一个新的翼螺旋因子WIN。北方印迹分析表明，WIN在多种胰岛素瘤细胞系和大鼠胚胎胰腺和肝脏中高度表达，在成年人中，WIN在胸腺、睾丸、肺和几个肠区域中检测到表达。发现WIN以高亲和力结合选择的序列5 ′-AGATTGAGTA-3 ′，其类似于最近鉴定的HNF-6结合序列5 ′-DHWATTGAYTWWD-3 ′。（其中W = A或T，Y = T或C，H不是G，D不是C），我们通过文库筛选和cDNA末端的5 ′-快速扩增分离了人WIN cDNA，序列分析表明人WIN的羧基末端先前已被分离为推定的磷酸化底物，MPM 2-反应性磷蛋白2（MPP 2）;大鼠和人WIN cDNA的比对以及与小鼠基因组序列的比较表明，WIN DNA结合结构域由四个外显子编码，其中两个外显子是由两个外显子编码的，其中两个外显子是由两个外显子编码的。（外显子4和6）被选择性剪接以产生至少三类mRNA转录物，RNase保护试验表明这些转录本在不同组织中有差异表达，翼螺旋DNA结合区的选择性剪接可能导致DNA结合特异性的调节。
We have cloned a novel winged helix factor, WIN, from the rat insulinoma cell line, INS-1. Northern blot analysis demonstrated that WIN is highly expressed in a variety of insulinoma cell lines and rat embryonic pancreas and liver, In adults, WIN expression was detected in thymus, testis, lung, and several intestinal regions, We determined the DNA sequences bound in vitro by baculovirus-expressed WIN protein ire a polymerase chain reaction-based selection procedure, WIN was found to bind with high affinity to the selected sequence 5'-AGATTGAGTA-3', which is similar to the recently identified HNF-6 binding sequence 5'-DHWATTGAYTWWD-3' (where W = A or T, Y = T or C, H is not G, and D is not C), We have isolated human WIN cDNAs by library screening and 5'-rapid amplification of cDNA ends, Sequence analysis indicates that the carboxyl terminus of human WIN has been previously isolated as a putative phosphorylation substrate, MPM2-reactive phosphoprotein 2 (MPP2); WIN may be regulated by phosphorylation, Alignment of the rat and human WIN cDNAs and their comparison with mouse genomic sequence revealed that the WIN DNA binding domain is encoded by four exons, two of which (exons 4 and 6) are alternatively spliced to generate at least three classes of mRNA transcripts, These transcripts were shown by RNase protection assay to be differentially expressed in different tissues, Alternative splicing within the winged helix DNA binding domain might result in modulation of DNA binding specificity.