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基4-基化调节阿尔戈诺特2的稳定性
Hank H Qi1, Pat P Ongusaha, Johanna Myllyharju
1Department of Pathology, Harvard Medical School, New Research Building 854, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA.
Nature
|August 12, 2008
概括
通过原蛋白prolyl-4-hydroxylase对Argonaute 2 (Ago2) 的氧化调节其稳定性. 这种翻译后的修改对于有效的RNA干扰和短干扰RNA活性至关重要.
科学领域:
- 分子生物学分子生物学
- 翻译后修改 翻译后修改
- 在RNA干扰过程中,RNA干扰
背景情况:
- 人类阿尔戈诺特 (Ago) 蛋白质是RNA诱导沉默复合体 (RISC) 的关键组成部分.
- Ago2的PIWI域介导了向RNA裂变,这是RNA干扰中的关键步骤.
- RISC的组装和功能涉及许多相关蛋白质,但调控机制尚不清楚.
研究的目的:
- 研究RNA干扰的新型调节机制.
- 为了确定Ago蛋白和其他细胞因子之间的相互作用.
- 阐明翻译后修改在Ago2稳定性和功能中的作用.
主要方法:
- 质谱测量以确定蛋白质相互作用和翻译后修改.
- 共同免疫沉以确认物理相互作用.
- 短发针RNA (shRNA) 和CRISPR/Cas9基因编辑以耗尽或禁用特定蛋白质.
- 在修饰细胞中分析Ago2稳定性和RNA干扰活性.
主要成果:
- 在Ago2和I型原prolyl-4-hydroxylase (C-P4H(I)) 的子单元之间确定了物理相互作用.
- 发现内源性Ago2在proline 700 (P700) 中被化.
- 耗尽C-P4H(I) 子单元或Ago2突变,P700会破坏Ago2的稳定,并损害RISC活动.
结论:
- 通过C-P4H(I) 在P700的Ago2的基化是一种新的翻译后修饰.
- 这种基化对于保持Ago2稳定性至关重要.
- 在调节RNA干扰效率方面,ago2氧化起着至关重要的作用.
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