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PP2CBETA的5种同工型中，在睾丸和肠道中明显表达了3种同工型，β -3，-4和-5。最近有关于pp2cbeta cDNA的5'末端的报道，分别为A型和B. B. slso，启动子（P1），该启动子（P1）最近在A型区域的5'the中揭示了无处不在的基因表达。为了研究pp2cbeta mRNA的5端的可能变化，进行了5'-race和该区域基因的克隆，并确定了3个外显子：A型AS exon1，b型A型AS型外显子3和外显子2作为新颖的外显子。 RT-PCR和Southern印迹分析表明，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'区域进一步研究为潜在的器官特异性启动子（P2）。通过底漆扩展实验观察到了多个转录起始位点，观察到了C-MOS共有序列的几个SRY和NRE，这意味着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测定法证明了P2的显着启动子活性。

项目成果

Yokoyama, N., Kobayashi, T., Tamura, S.& Sugiya, H.: "Purification and characterization of protein phosphatase 2C in rat parotid acinar cells : two forms of Mg^<2+>-activated histone phosphatase and phosphorylation by cAMP-dependent protein kinase." Arch.

横山，N.，小林，T.，田村，S.

Ohnishi, M., Nakagawara, K., Mori, M., Kobayashi, T., Kato, S., Sasahara, Y., Kusuda, K., Chida, N., Kobayashi, T., Yanagawa, Y., Hiraga, A., Takeuchi, T.& Tamura, S.: "Localization of the Mouse protein serine/threonine phosphatase 2Cbeta gene to chromoso

大西，M.，中河原，K.，森，M.，小林，T.，加藤，S.，笹原，Y.，楠田，K.，千田，N.，小林，T.，柳川，Y.，

Shunsuke, Kato: "Differentiation-dependent enhanced expression of phosphatase 2Cβ in germ cells of mouse seminiferous tubules." FEBS Lett.396. 293-297 (1996)

Shunsuke, Kato：“小鼠生精小管生殖细胞中磷酸酶 2Cβ 的分化依赖性增强表达。”FEBS Lett.393-297 (1996)。

Yick Pang Ching: "Specificity of different isoforms of protein phosphatase 2A and protein phosphatase 2C studied using site-directed mutagenesis of HMG-CoA reductase." FEBS Lett.411. 265-268 (1997)

Yick Pang Ching：“利用 HMG-CoA 还原酶的定点诱变研究了蛋白磷酸酶 2A 和蛋白磷酸酶 2C 不同亚型的特异性。”

Takayasu Kobayashi: "Enhanced UV sensitivity of yeast cells induced by over expression of Mg^<2+>-dependent protein phosphatase α (Type 2Cα)。" Mutation Res.362. 213-217 (1996)

Takayasu Kobayashi：“Mg^2+依赖性蛋白磷酸酶α（2Cα型）的过度表达诱导酵母细胞的紫外线敏感性增强。”213-217（1996）。