Molecular mchanisms of stress tolerance observed at mildly low temperature environment and the clinical relevance of their abnormality in the testis

微低温环境下睾丸应激耐受的分子机制及其异常的临床意义

• 批准号: 18390434
• 负责人: FUJITA Jun
• 金额: $ 10.17万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18390434/
• 关键词: hypothermia; environment; testis; transcription factor; stress; 遺伝子; 細胞・組織; 特殊環境; cirp

In the mammalian testes, germ cells and somatic colic undergo active cell proliferation and differentiation at mildly low temperatures (32-34℃). To elucidate how testicular cells adapt to the lower temperature and become more resistant to various stressors, we have analyzed the molecular mechanism underlying the cold shock response, especially in relation to the cold shock protein alp (cold-inducible RNA-binding protein).1. We failed to identify the transcription factor that binds to the mild-cold response element (MCRE) and activates transcription at lower temperatures by the yeast 1-hybrid screening, but successfully identified several proteins by mass spectrometry that associate with MORE in vitro and in vivo. Most of them seemed to modify activity of the elusive transcription factor.2. When over-expressed in cultured rolls, several of the identified "MCRE-binding proteins" enhanced the expression of Cap as well as the reporter gene driven by MCRE and CMV promoter One of the genes was transiently over-expressed in cultured cells and the induced genes similarly to Cirp were identified by using a DNA microarray.3. MCRE was found to be ineffective as a cold-responsive enhancer when it is integrated into the chromosome at a cm all copy numbers. We optimized the sequence and copy numbers for better induction at 32℃.4. We produced the dip knockout mice and demonstrated that Cirp is involved in wound healing process.5. By using the yeast 2-hybrid screening, we identified proteins, including a kinase, that interact with Cirp and potentially endow resistance to apoptotic stresses.

在哺乳动物睾丸中，生殖细胞和体细胞结肠在轻度低温（32-34℃）下进行活跃的细胞增殖和分化。为了阐明睾丸细胞如何适应低温并对各种应激源更具抵抗力，我们分析了冷休克反应的分子机制，特别是与冷休克蛋白alp（冷诱导RNA结合蛋白）的关系。通过酵母单杂交筛选，我们未能鉴定出与微冷反应元件（MCRE）结合并在较低温度下激活转录的转录因子，但通过质谱法成功地鉴定出与MORE在体外和体内相关的几种蛋白。其中大多数似乎改变了难以捉摸的转录因子的活性.当在培养卷中过表达时，几个鉴定的“MCRE结合蛋白”增强Cap以及由MCRE和CMV启动子驱动的报告基因的表达。其中一个基因在培养细胞中瞬时过表达，并且通过使用DNA微阵列鉴定类似于Cirp的诱导基因。MCRE被发现是无效的冷响应增强子时，它是整合到染色体中的cm所有拷贝数。我们优化了序列和拷贝数，以便在32℃下更好地诱导.我们建立了dip基因敲除小鼠模型，证明了Cirp参与了创伤愈合过程.通过使用酵母双杂交筛选，我们鉴定了与Cirp相互作用并可能赋予对凋亡应激的抗性的蛋白质，包括激酶。

项目成果

Regulatory mechanisms that specificaly enhance gene expression at mildly low temperature in the testis

在睾丸中轻度低温下特异性增强基因表达的调节机制

• 发表时间: 2007
• 作者: Jun;Fujita
• 通讯作者: Fujita

Enhanced deacetylation of p53 by the anti-apoptotic protein HSCO in association with HDAC1.

与 HDAC1 相关的抗凋亡蛋白 HSCO 增强了 p53 的脱乙酰作用。

• 发表时间: 2007
• 期刊: J Biol Chem (印刷中)
• 作者: Valcourt U;Kowanetz M;Niimi H;Heldin CH;Moustakas A;北島 勲;北島 勲;Higashitsuji H
• 通讯作者: Higashitsuji H

哺乳類低温ショック蛋白質Cirpの示す抗アポトーシス作用

哺乳动物冷休克蛋白 Cirp 的抗凋亡作用

• 发表时间: 2006
• 作者: Jun;Fujita;藤田 潤
• 通讯作者: 藤田 潤

Cirp protects against tumor necrosis factor-alpha-induced apoptosis via activation of extracellular

Cirp 通过激活细胞外细胞来防止肿瘤坏死因子-α 诱导的细胞凋亡

• 发表时间: 2006
• 期刊: Biochim Biophys Acta 1763
• 作者: Valcourt U;Kowanetz M;Niimi H;Heldin CH;Moustakas A;北島 勲;北島 勲;Higashitsuji H;Dawson S;Mayer RJ;Yamaguchi K;Sakurai T
• 通讯作者: Sakurai T

軽度の低温により遺伝子の発現を促進させる配列

轻度低温促进基因表达的序列

• 发表时间: 2010