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，3 种亚型（β-3、-4 和 -5）在睾丸和肠道中独特表达。已有关于 PP2Cbeta cDNA 5' 端变异的报道，指定为 A 型和 B 型。此外，最近在 A 型区域的 5' 端发现了一个启动子 (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，对该区域的基因进行了5'-RACE和克隆，并鉴定了3个外显子：A型为外显子1，B型为外显子3，外显子2为新外显子。 RT-PCR和Southern印迹分析显示外显子2仅在睾丸、肠和肝脏中表达。基于这些结果，外显子 2 的 5' 区域被进一步研究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，并观察到多个c-mos共有序列的SRY和NRE，提示P2可能参与性分化和睾丸功能。 P19 EC 细胞的荧光素酶报告基因检测证明了 P2 显着的启动子活性。

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