Regulation of PP2Cbeta gene expression-testis specific expression and multiple promoter

PP2Cbeta基因表达的调控——睾丸特异性表达和多启动子

• 批准号: 08670133
• 负责人: TAMURA Shinri
• 金额: $ 1.41万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670133/
• 关键词: protein phosphatase 2Cbeta; genomic DNA; promoter; testis; プロモーター; プロテインホスファターゼ; 遺伝子発現; 精巣

Out of known 5 isoforms of PP2Cbeta, 3 isoforms, beta-3, -4 and -5, are distinctively expressed in testis and intestine. There has been reports on the variation of 5' end of the PP2Cbeta cDNAs, assigned as type A and B.Also, a promoter (P1), which enables ubiquitous expression to the gene, was recently brought to light at 5' of type A region. In order to investigate the possible variations of the 5' end of the PP2Cbeta mRNAs, 5'-RACE and cloning of the gene of that region were performed, and 3 exons were identified : type A as exon1, type B as exon 3 and exon 2 as a novel one. RT-PCR and Southern blotting analysis showed the exon 2 is expressed only in testis, intestine and liver. Based on these results, 5' region of the exon 2 was further investigated as a potential organ-specific promoter (P2). Multiple transcription start sites were observed by the primer extention experiment, and several SRY and NRE of c-mos consensus sequences were observed, implying the possible involvement of the P2 in sexual differentiation and testicular function. Luciferase reporter assay with P19 EC cells proved significant promoter activity of P2.

在已知的5种PP2Cbeta亚型中，β -3、-4和-5亚型在睾丸和肠道中特异性表达。已经有关于pp2beta cdna 5‘端变异的报道，被分配为A型和b型。此外，最近在A型区域的5’处发现了一个启动子（P1），该启动子使该基因能够普遍表达。为了研究PP2Cbeta mrna 5′端可能的变异，我们对该区域的5′-RACE基因进行克隆，鉴定出3个外显子：A型为外显子1，B型为外显子3,2为新外显子。RT-PCR和Southern blotting分析显示，外显子2仅在睾丸、肠道和肝脏中表达。基于这些结果，我们进一步研究了外显子2的5'区域作为潜在的器官特异性启动子（P2）。引物延伸实验观察到多个转录起始位点，并观察到c-mos一致序列的多个SRY和NRE，暗示P2可能参与性别分化和睾丸功能。P19 EC细胞荧光素酶报告试验证实P2启动子活性显著。

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