Mechanism of The Activation of 2'-5'Oligoadenylate Synthetase by Double-Stranded RNA

双链RNA激活2-5寡腺苷酸合成酶的机制

• 批准号: 05680548
• 负责人: SOKAWA Yoshihiro
• 金额: $ 1.34万
• 依托单位: KYOTO INSTITUTE OF TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05680548/
• 关键词: 2'-5'OLIGOADENYLATE SYNTHETASE; INTERFERON; DOUBLE-STRANDED RNA; ウエスタン解析; ノーザン解析; 免疫組織化学; 2´-5´オリゴアデニル酸合成酵素; インターフェロン誘導酵素; 2-5A合成酵素

1. Nature of mouse 2'-5'oligoadenylate synthetase (2-5AS)The ORF of mouse 2-5AScDNA encodes 367 amino acid residues (42kD) , and two subtypes (S^+ and S^-) of the cDNA have been isolated, which are different in 14 amino acids. The enzymatic activity of S^- is only about 1/1000 of that of S^+, while the S^-mutant, whose 176Trp is converted to 176Glu, has an increase of activity by 30 times of that of S^-. The S^+ enzyme had a higher binding activity to double stranded RNA comparing with S^-, but the S^- mutant had a similar activity to S^+. These results indicate that the structure of the central part of the enzyme is responsible both to the enzyme activity and to the binding to double stranded RNA.2. Physiological expression of the 2-5AS gene in mouse intestineThe normal mouse, not treated exogenously with any IFN of IFN inducers, has been shown to have an enhanced level of 2-5AS activity. Among the organs of normal mouse, intestinal tissues had the highest levels of 2-5AS mRNA.Furthermore, the 42kD 2-5AS mRNA was expressed in intestines from germ-free mice and fetuses. Immunoblotting analysis using a monoclonal antibody against the 42kD 2-5AS revealed that the 42kD enzyme as well as a cross-reactive protein of 30kD were produced in the organs of normal mouse.

1.小鼠2'-5'寡腺苷酸合成酶(2-5AS)的性质小鼠2-5AScDNA的ORF编码367个氨基酸残基(42kD)，已分离出该cDNA的两个亚型(S^+和S^-)，它们有14个氨基酸不同。 S^-的酶活性仅为S^+的1/1000左右，而176Trp转化为176Glu的S^-突变体，其酶活性比S^-提高了30倍。与S^-相比，S^+酶对双链RNA具有更高的结合活性，但S^-突变体具有与S^+相似的活性。这些结果表明酶的中心部分的结构对酶活性和与双链RNA的结合都有影响。2。小鼠肠道中2-5AS基因的生理表达未用任何IFN或IFN诱导剂进行外源处理的正常小鼠已显示出具有增强水平的2-5AS活性。在正常小鼠的器官中，肠道组织中2-5AS mRNA的含量最高。此外，42kD的2-5AS mRNA在无菌小鼠和胎儿的肠道中也有表达。使用针对 42kD 2-5AS 的单克隆抗体进行的免疫印迹分析表明，正常小鼠的器官中产生了 42kD 酶以及 30kD 的交叉反应蛋白。

项目成果

宗川吉汪: "Physiological Expression of the 2´,5´-Oligoadenylate Synthetase Gene in Mouse Intestine" Journal of Interferon Research. 14. 121-127 (1994)

Yoshitaka Munekawa：“小鼠肠道中 2´,5´-寡腺苷酸合成酶基因的生理表达”干扰素研究杂志 14. 121-127 (1994)。

YOSHIHIRO SOKAWA: "Physiological Expression of 2', 5'-Oligoadenylate Synthetase Gene in Mouse Intestine" Journal Interferon Research. Vol.14. 121-127 (1994)

YOSHIHIRO SOKAWA：“小鼠肠道中 2, 5-寡腺苷酸合成酶基因的生理表达”干扰素研究杂志。

宗川吉汪: "Physiolgical Expression of the 2'5'-Oligoadenylate Synthetase Gene in Mouse Intestine" Journal of Interferon Research. 14. 121-127 (1994)

Yoshitaka Munekawa：“小鼠肠道中 25-寡腺苷酸合成酶基因的生理表达”干扰素研究杂志 14. 121-127 (1994)。

伊勢川裕二: "Strand-Specific Detection of Hantaan Viyus RNA Sequences by In Vifro DNA Amplification" Microbiology and Immunology. 38. 905-908 (1994)

Yuji Isekawa：“通过体外 DNA 扩增对 Hantaan Viyus RNA 序列进行链特异性检测”微生物学和免疫学 38. 905-908 (1994)。

YUJI ISEGAWA: "Strand-Specific Detection of Hantaan Virus RNA Sequences by In Vitro DNA Amplification" Microbiology and Immunology. Vol.38. 905-908 (1994)

YUJI ISEGAWA：“通过体外 DNA 扩增对汉滩病毒 RNA 序列进行链特异性检测”微生物学和免疫学。