与亚结合的T7促进剂,用于高效的转录光调节
Mingzhe Liu1, Hiroyuki Asanuma, Makoto Komiyama
1Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Journal of the American Chemical Society
|January 19, 2006
概括
研究人员在T7促进体中使用亚博烯开发了可光控制的转录. 引入亚博烯允许光调节T7RNA聚合酶 (T7RNAP) 活动,提高转录效率.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 合成生物学 合成生物学
背景情况:
- 用外部刺激控制基因转录对于合成生物学应用至关重要.
- 光调节可以对生物过程提供精确的时间和空间控制.
- T7促进体是用于高水平基因表达的广泛使用的系统.
研究的目的:
- 通过使用亚博烯,对T7促进体进行光控制的转录.
- 为了研究亚博烯位置对T7RNA聚合酶 (T7RNAP) 活性的影响.
- 通过多个亚博烯修饰的协同效应来实现增强的光控制.
主要方法:
- 在特定区域 (-7至-11和1-1至-4) 用亚博烯对T7促进体进行化学修饰.
- 紫外线照射以诱导亚博的 cis-异体化和控制转录.
- 动力分析 (Km和kcat的确定),以阐明光调节的机制.
主要成果:
- 单次亚博引入抑制了转化形式的转录,并增强了 cis 形式的转录 (1.5-2.0 倍的增加).
- 在循环结合区域的亚烯影响了Km,而在解区域的亚烯影响了kcat.
- 在不同区域的双双阿佐基修饰在紫外线照射时产生了协同作用的7.6倍的转录增加.
结论:
- 加入阿佐为T7RNAP转录提供了有效的光控制.
- 亚博烯的位置对转录调节的动态参数具有关键的影响.
- 在特定的促进区域内,两种亚的合作行动可以实现强大的和增强的光调节.
相关概念视频
Transfer RNA Synthesis
One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases.
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
tRNA Activation
Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
Master Transcription Regulators
Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...
Transcription Attenuation in Prokaryotes
Transcriptional attenuation occurs when RNA transcription is prematurely terminated due to the formation of a terminator mRNA hairpin structure. Bacteria use these hairpins to regulate the transcription process and control the synthesis of several amino acids including histidine, lysine, threonine, and phenylalanine. Transcription attenuation takes place in the non-coding regions of mRNA.
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
There are several different mechanisms used to attenuate transcription. In ribosome mediated...
tRNA Activation
Aminoacyl-tRNA synthetases are present in both eukaryotes and bacteria. Though eukaryotes have 20 different aminoacyl-tRNA synthetases to couple to 20 amino acids, many bacteria do not have genes for all of these aminoacyl-tRNA synthetases. Despite this, they still use all 20 amino acids to synthesize their proteins. For instance, some bacteria do not have the gene encoding the enzyme that couples glutamine with its partner tRNA. In these organisms, one enzyme adds glutamic acid to all of the...
Master Transcription Regulators
Master transcription regulators are regulatory proteins that are predominantly responsible for regulating the expression of multiple genes. Often these genes work in concert to drive a complex process. Activation of a master transcription regulator can lead to a cascade of transcriptional activation necessary for that outcome. These regulators can directly bind to the regulatory sequences of the various genes involved, or they can indirectly regulate transcription by binding to regulatory...


