Research for the mechanism of mRNA degradation in animal cells.

动物细胞mRNA降解机制研究。

• 批准号: 06672167
• 负责人: HIROSE Seiyu
• 金额: $ 1.34万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06672167/
• 关键词: RAT LIVER; RIBONUCLEASE; PDG-A; AORTIC SMC; 5'-UTR; uORF; mRNA STABILITY; SUBSTRATE SPECIFICITY; 5′非翻訳領域; 核酸結合タンパク質; mRNA分解; 分子生物学

PURIFICATION OF RIBONUCLEASES (HIROSHI KUMAGAI). The novel RNase were purified from rat liver cytosomal fraction ; molecular weight 13 kDa and 16 kDa RNases were termed RNase 13 and RNase 16, respectively. RNase 16 hydrolyzed in the order of poly (C) >yeast total RNA >poly (U) and may be inbolved amember of pancreatic RNase family. On the other hand, RNase 13 hyodrolyzed in the order of poly (U) >yeast total RNA >Poly (C) and may be inbolved a member of plasma RNase family.POSSIBLE RELATIONSHIP OF THE STABILTY OF mRNA AND ACTIVITY OF A SITE SPECIFIC RNase. (TOMOHITO KAKEGAA). Platelet derived growth factor (PDGF) A plays an important role in the proliferation and chemostasis of animal cells. The most suitable candidate of region of PDGF-A mRNA for site specific degradation was investigated in rat cultured smooth muscle cells. Ultra violet derived crosslinking reaction and RNase protection assay revealed the terminal codon area of the upper open reading frame existing in the 5' untranslated region (5'-UTR) PDGF-A had high potential for the start site of this mRNA.Four kinds of purified rat liver RNases except exo-RNase that was purified from rat liver microsomal fraction did not degrade site specifically with PDGF-A 5'-UTR.This results suggest that site specific degradation of PDGF-A 5'-UTR by three endo-RNases requires additional unknown endogenous factors. The new application of the North-western blot for detection of RNase activity revealed that cytosomal fraction contained many logical candidates for the sequence specific RNase activity. This result suggest that the application of North-western blot for the detection of RNase activity may be useful for determination and isolation of site specific RNA degrading activity.

核糖核酸酶的纯化（kumagai）。从大鼠肝脏胞体级分纯化了新型的RNase；分子量13 kDa和16 kDa RNass分别称为RNase 13和RNase 16。 RNase 16按聚（c）>酵母总RNA> poly（u）的顺序水解，并且可以在胰腺RNase家族的为为为象征的室。另一方面，rNase 13按聚（i）>酵母总RNA> poly（c）的顺序hyodropy，并且可以在为叶等血浆rnase家族的成员中。 。 （Tomohito Kakegaa）。血小板衍生的生长因子（PDGF）A在动物细胞的增殖和化学上起着重要作用。在大鼠培养的平滑肌细胞中研究了最合适的PDGF-A mRNA区域mRNA候选者。超紫衍生的交联反应和RNase保护测定法揭示了5'未翻译区域（5'-utr）PDGF-A在此mRNA的起始地点的潜力很高。4纯化的大鼠肝脏RNase除外，除了从大鼠肝微粒体级分纯化的外脱酶没有特异性降解位点，这表明PDGF-A 5'-UTR的位点特异性降解被三个内部范围降解。需要其他未知的内源性因素。西北印迹在发现RNase活性中的新应用显示，胞体分数包含许多序列特异性RNase活性的逻辑候选。该结果表明，西北印迹在检测RNase活性中的应用可能有助于确定和分离特定的RNA降解活性。