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High-throughput Purification of Affinity-tagged Recombinant Proteins
Published on: August 26, 2012
转录因子IIB对自己进行乙化,以调节转录
Chu H Choi1, Makoto Hiromura, Anny Usheva
1Endocrinology Division, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 99 Brookline Avenue, Boston, Massachusetts 02215, USA.
Nature
|August 22, 2003
概括
一般转录因子TFIIB经过乙化,是一种翻译后的修改. 这种自化稳定了相互作用并激活了基底转录,将乙-CoA与基因调节联系起来.
科学领域:
- 分子生物学分子生物学
- 基因规则 基因规则
- 生物化学 生物化学
背景情况:
- 乙化是一种已知的调节后翻译性修改.
- 乙化在调节基底转录因子中的作用在很大程度上仍未被探索.
- 一般的转录因子对于启动真核转录至关重要.
研究的目的:
- 研究乙化在基底转录因子调节中的作用.
- 要确定TFIIB (一般转录因子II B) 是否受到乙化.
- 阐明TFIIB乙化的功能后果.
主要方法:
- 生物化学分析检测和描述TFIIB乙化.
- 在体外转录分析使用复合和细胞TFIIB.
- 局部定向突变生成以产生非可乙化TFIIB突变体 (K238A).
- 对TFIIB-TFIIF互动的分析.
主要成果:
- TFIIB在特定的氨酸残留物 (K238) 中被乙化.
- TFIIB具有自身乙转移酶活性,利用乙-CoA.
- 乙化稳定了TFIIB和TFIIF之间的相互作用.
- TFIIB自化增强基底转录启动在体外和细胞内.
- 一种非可乙化TFIIB的K238A突变体未能激活转录.
结论:
- TFIIB乙化是一种基础转录的新型调节机制.
- TFIIB作为一种自身乙转移酶,将乙-CoA与转录联系起来.
- TFIIB的乙化在稳定转录因子相互作用和促进基因表达方面发挥着关键作用.
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