Analysis on biological formation of a novel modified nucleic acid, 8-nitorguanosine, and its unique signal function

新型修饰核酸8-硝基鸟苷的生物形成及其独特的信号功能分析

• 批准号: 16590249
• 负责人: AKUTA Teruo
• 金额: $ 2.18万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590249/
• 关键词: NO; 8-nitroguanosine; cGMP; 8-nitroc-GMP; anti-8-nitro-cGMP antibody; protein kinase G; cell signaling; redox signaling; 抗8-ニトログアノシン抗体; 細胞保護作用; ヘムオキシゲナーゼ-1; RNAウイルス; 遺伝子変異

We examined biological and signal function of a novel nucleic acids modification by NO, especially, 8-nitroguanosine (8-NO_2Guo) and 8-NO_2Guo-related compounds, e.g. 8-nitro-cyclic GMP (8-NO_2-cGMP). Our data on these compounds are summarized as follows.1) 8-NO_2Guo formation was mainly localized in cytosol, near the endoplasmic reticulum, in HepG2 cells treated with NO, as identified by confocal microscopy and immunoelectron microscopy analysis using anti 8-NO_2Guo monoclonal antibody. We demonstrated that 8-NO_2Guo protected from apoptosis HepG2 cell through upregulation of heme oxygenase-1 and endothelial NOS (eNOS), and inhibition of bcl-XL degradation.2) 8-NO_2Guo has mutagenic potential in DNA and RNA. Authentic 8-NO_2Guo added exogenously to cultured CV-1 cells infected with a recombinant Sendai virus containing a green fluorescent protein gene increased the viral mutation frequency. This mutation spectrum was dominantly observed with a C to U transition. 8-NO_2Guo added to AS5 … More 2 cells also increased the genomic mutation frequency, with a predominant mutation, G to T transition. We supposed two different mechanisms for this mutagenesis : direct modification of nucleic acid and indirect augmentation of oxidative stress via superoxide generation by 8-NO_2Guo.3) We established the synthesis method of 8-NO_2-cGMP and prepared large amount of this compound. Treatment of human uterine smooth muscle cells with 8-NO_2-cGMP increased in cGMP-dependent protein kinase-associated phosphorylation of vasodilator-stimulated phosphoprotein on Ser^<239>. Strong vasorelaxing potential for rat carotid artery was observed with 8-NO_2-cGMP. These results suggested that 8-NO_2-cGMP is a new cellular redox signaling molecule.4) We developed the anti-8-NO_2-cGMP monoclonal and polyclonal antibodies. The immunostaining with anti-8-NO_2-cGMP monoclonal antibody showed high immuoreactivity in cytosol of mouse macrophage derived RAW264 cells stimulated with IFN-γ and lipopolysaccharide.Taken together, 8-NO_2Guo and 8-NO_2-cGMP have unique biochemical and pharmacological properties such as cellular redox signaling and mutagenic potential. Less

我们研究了一种新型核酸修饰的生物学和信号功能，特别是8-硝基鸟苷（8-NO_2Guo）和8-NO_2Guo相关化合物，如8-硝基环GMP （8-NO_2-cGMP）。我们对这些化合物的数据总结如下。1)利用抗8-NO_2Guo单克隆抗体共聚焦显微镜和免疫电镜分析发现，NO作用的HepG2细胞中，8-NO_2Guo的形成主要集中在靠近内质网的细胞质中。我们发现8-NO_2Guo通过上调血红素加氧酶-1和内皮细胞NOS (eNOS)，抑制bcl-XL降解来保护HepG2细胞免于凋亡。2) 8-NO_2Guo对DNA和RNA具有诱变潜力。8-NO_2Guo原液外源性添加到含有绿色荧光蛋白基因的重组仙台病毒感染的培养CV-1细胞中，增加了病毒的突变频率。该突变谱以C向U的转变为主。在AS5中加入8-NO_2Guo，增加2个细胞也增加了基因组突变频率，以G向T转变为优势突变。我们推测这种诱变的两种不同机制：直接修饰核酸和通过8-NO_2Guo产生超氧化物间接增强氧化应激。3)建立了8-NO_2-cGMP的合成方法，并大量制备了该化合物。8-NO_2-cGMP处理人子宫平滑肌细胞后，cgmp依赖性蛋白激酶相关的Ser^<239>血管扩张剂刺激磷酸化增加。8-NO_2-cGMP观察到大鼠颈动脉有较强的血管舒张电位。这些结果表明8-NO_2-cGMP是一种新的细胞氧化还原信号分子。4)制备抗8- no_2 - cgmp单克隆和多克隆抗体。抗8- no_2 - cgmp单克隆抗体免疫染色显示，IFN-γ和脂多糖刺激的小鼠巨噬细胞源性RAW264细胞胞浆具有较高的免疫反应性。综上所述，8-NO_2Guo和8-NO_2-cGMP具有独特的生化和药理学特性，如细胞氧化还原信号传导和致突变潜力。少

项目成果

Antioxidative and antimutagenic activities of 4-vinyl-2,6-dimethoxyphenol (Canolol) isolated from canola oil

• DOI: 10.1021/jf040045
• 发表时间: 2004-07-14
• 期刊: JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
• 影响因子: 6.1
• 作者: Kuwahara, H;Kanazawa, A;Maeda, H
• 通讯作者: Maeda, H

Interleukin-1beta induces death in chondrocyte-like ATDC5 cells through mitochondrial dysfunction and energy depletion in a reactive nitrogen and oxygen species-dependent manner.

• DOI: 10.1042/bj20041996
• 发表时间: 2005-07
• 期刊: The Biochemical journal
• 作者: R. Yasuhara;Y. Miyamoto;T. Akaike;T. Akuta;Masanori Nakamura;M. Takami;N. Morimura;Kayoko Yasu;R. Kamijo

Nitric oxide-induced nitrative stress involeved in microbial pathogenesis.

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• 发表时间: 2005
• 期刊: J. Pharmacol. Sci. 8
• 作者: Zaki;M.H.et al.
• 通讯作者: M.H.et al.

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一氧化氮气体杀死病原体：一氧化氮对生物分子的硝化和亚硝化作用介导的独特抗菌作用

• 发表时间: 2006
• 期刊: 現代科学 424(印刷中)
• 作者: 佐伯武頼;小林圭子;堀内正久(分担);澤 智裕 他
• 通讯作者: 澤 智裕 他

Quantitative assessment of protein-bound tyrosine nitration in airway secretions from patients with inflammatory airway diseases.

炎症性气道疾病患者气道分泌物中蛋白质结合酪氨酸硝化的定量评估。

• 发表时间: 2004
• 期刊: Free Rad. Res. 38
• 作者: Sugiura;H. et al.
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