PRR11（proline-rich protein 11，PRR11）是我们最近发现的一个新的肿 瘤相关基因.初步研究表明, PRR11参与细胞增殖、细胞周期和细胞癌变等多种生 物学过程．为了进一步研究PRR11基因的转录调控机制并全面解析其功能，本研究 对PRR11基因的启动子进行了克隆鉴定和初步分析．首先,应用5' RACE（rapid amplification of cDNA ends，cDNA末端快速扩增）技术鉴定了PRR 11基因的转 录起始位点，发现了其具有多个转录起始位点．通过PCR定向克隆和DNA blunting 技术，构建了6个相互重叠并覆盖PRR 11基因转录起始位点附近约2.0 kb区域的 PRR 11基因启动子荧光素酶报告基因重组体．启动子活性分析表明，PRR 11基因 启动子主要定位于转录起始位点附近-563 bp～+341 bp的区域内．采用转录因子 结合位点预测分析软件分析表明，PRR 11基因启动子缺乏典型的TATA盒，但含有 典型的GC盒、CCAAT盒以及潜在的经典转录因子E2F1和MYB的结合位点，提示Sp1、 NF-Y、E2F1和MYB等经典转录因子可能参与PRR 11基因的转录调控．
Abstract
Proline-rich protein 11(PRR11) is a newly discovered tumor-related gene. PRR11 plays an important role in cell proliferation, cell cycle and carcinogenesis. To further investigate its transcriptional regulatory mechanism and biological function, PRR11 gene promoter was cloned and identified in the present study. At first, the transcriptional start sites for PRR11 gene were identified by 5' RACE (rapid amplification of cDNA ends) and bioinformatic analysis. 5' RACE assay indicated that PRR11 gene has multiple distinct transcriptional start sites. Furthermore, several overlapping genomic fragments from the 5'-flanking region of PRR11 gene were coloned into pGL3-basic vector to construct PRR11 promoter reporters. Luciferase reporter assay indicated that PRR11 promoter region is mainly located in a region of -563 bp to +341 bp around the major transcriptional start site. Transcriptional factor binding analysis indicated that PRR11 gene promoter lacks of classical TATA box, but contains classical GC box, CCAAT box, as well as other putative binding sites for classic transcriptional factors such as E2F1 and MYB. These results suggested that transcriptional factors such as Sp1, NF-Y, E2F1 and MYB might be involved in the transcriptional regulation of PRR11 gene.
关键词
PRR11（proline-rich protein 11) /
启动子 /
转录调控 /
肿瘤相关基因
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Key words
proline-rich protein 11gene (PRR11) /
promote /
r transcriptional regulation /
tumor-related gene
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中图分类号：
Q78
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脚注
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基金
国家自然科学基金（No.30801356, No.81001097）,重庆市科委自然科学基金（No.2010BB5357）和重庆市教委科学技术研究项目（No.KJ090305）资助
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