Okazaki M, Ito S, Kawakita K, Takeshita S, Kawai S, Makishima F, Oda H, Kakinuma A
Discovery Research Laboratories, Hoechst Marion Roussel Ltd., Kawagoe, 350-1162, Japan.
Genomics. 1999 Aug 15;60(1):87-95. doi: 10.1006/geno.1999.5902.
The preferential screening of cDNA libraries derived from the mouse osteoblastic cell line MC3T3-E1 has yielded a cDNA clone encoding a 442-amino-acid protein designated STAP (signal transduction and adaptor protein), which contains several motifs shared among transcription factors and adaptors such as a Zn-finger like motif, a proline-rich domain, and a PEST sequence. The amino acid sequence homology search also reveals that STAP is identical to a mouse oxidative stress protein, A170, and has 90% homology with a human p62 protein that binds to the tyrosine kinase p56(lck) SH2 domain. Northern blot analysis indicated a broad expression profile of STAP mRNA in various tissues and cell lines. In MC3T3-E1 cells, STAP mRNA was induced by treatment with TGF-beta, but not with BMP-2 or GDF-5. Analysis of the mouse STAP gene isolated from the genomic library revealed that the STAP gene spans a region of over 11 kb and comprises eight exons. The transcription start site was identified by primer extension analysis to be located 35 bp upstream from the translation initiation site. Sequencing analysis of the 5' flanking region of the STAP gene revealed multiple consensus motifs/sequences for several DNA binding transcription factors. The STAP gene had a TATA box, but no CCAAT box. Potential Sp1, AP-1, NF-E2, MyoD, and NF-kappaB binding sites were found in the 5' flanking region (1.4 kb) of the STAP gene.
对从小鼠成骨细胞系MC3T3-E1中获得的cDNA文库进行优先筛选，得到了一个编码442个氨基酸的蛋白质的cDNA克隆，该蛋白质被命名为STAP（信号转导和衔接蛋白），它包含转录因子和衔接蛋白中常见的几个基序，如锌指样基序、富含脯氨酸的结构域和PEST序列。氨基酸序列同源性搜索还显示，STAP与小鼠氧化应激蛋白A170相同，并且与一种与人酪氨酸激酶p56(lck) SH2结构域结合的人p62蛋白具有90%的同源性。Northern印迹分析表明，STAP mRNA在各种组织和细胞系中广泛表达。在MC3T3-E1细胞中，TGF-β处理可诱导STAP mRNA表达，但BMP-2或GDF-5处理则不能。对从基因组文库中分离出的小鼠STAP基因的分析表明，STAP基因跨越超过11 kb的区域，由八个外显子组成。通过引物延伸分析确定转录起始位点位于翻译起始位点上游35 bp处。对STAP基因5'侧翼区域的测序分析揭示了几种DNA结合转录因子的多个共有基序/序列。STAP基因有一个TATA盒，但没有CCAAT盒。在STAP基因的5'侧翼区域（1.4 kb）中发现了潜在的Sp1、AP-1、NF-E2、MyoD和NF-κB结合位点。
