Wang Chen-Yu, Liang Yuh-Jin, Lin Young-Sun, Shih Hsiu-Ming, Jou Yuh-Shan, Yu Winston C Y
Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 114, Taiwan.
J Biol Chem. 2004 Apr 23;279(17):17750-5. doi: 10.1074/jbc.M310532200. Epub 2004 Jan 26.
With the aim of identifying potential cellular proteins that mediate the transcriptional regulation of YY1, a HeLa cDNA library was screened using the yeast two-hybrid system. A previously unknown protein interacting with YY1 was identified and named YY1AP. By using the 5'-rapid amplification of cDNA ends technique, the full-length cDNA of YY1AP was cloned and sequenced. The cDNA was 2253 bp in length and encoded an open reading frame of 750 amino acids. The chromosomal gene was made up of 10 exons separated by nine introns and is localized on chromosome 1 (1q21.3). Northern blot analysis revealed that YY1AP is ubiquitously expressed in various human tissues and cancer cell lines. Co-immunoprecipitation and immunostaining of cells further indicated that YY1AP co-localizes with YY1 in the nucleus. Furthermore, YY1AP was shown to be capable of enhancing the transcriptional activation of an YY1 responsive promoter. Subsequent analysis by glutathione S-transferase pull-down assay showed that YY1AP contained two YY1 binding regions. The transactivation region of YY1AP would seem to be localized within the section of amino acids 260-345. It is proposed that YY1AP is a novel co-activator of YY1.
为了鉴定介导YY1转录调控的潜在细胞蛋白,利用酵母双杂交系统筛选了HeLa细胞cDNA文库。鉴定出一种与YY1相互作用的未知蛋白,并将其命名为YY1AP。通过使用5'-cDNA末端快速扩增技术,克隆并测序了YY1AP的全长cDNA。该cDNA长度为2253 bp,编码一个由750个氨基酸组成的开放阅读框。染色体基因由10个外显子和9个内含子组成,定位于1号染色体(1q21.3)。Northern印迹分析显示YY1AP在各种人类组织和癌细胞系中普遍表达。细胞的共免疫沉淀和免疫染色进一步表明YY1AP与YY1在细胞核中共定位。此外,YY1AP能够增强YY1反应性启动子的转录激活。随后通过谷胱甘肽S-转移酶下拉试验分析表明YY1AP包含两个YY1结合区域。YY1AP的反式激活区域似乎位于氨基酸260-345区段内。有人提出YY1AP是YY1的一种新型共激活因子。