使用光基 analogue 进行探测和扰乱带状交换机折叠
Janson E Hoeher1, Natalie E Sande1, Julia R Widom1
1Department of Chemistry and Biochemistry, University of Oregon, Eugene, Oregon, USA.
Photochemistry and photobiology
|December 15, 2023
概括
在 preQ1 рибо开关使用富含腺因的序列来结合连接体. 将光探针 (2-aminopurine) 插入这些序列中,会破坏连接体的结合,从而揭示了腺因在基因调节中的关键作用.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 在RNA生物学,RNA生物学.
背景情况:
- 丝带切换器是调控mRNA元素,控制基因表达.
- 一个I类preQ1核突变器在结合联体后形成一个伪结结构.
- 在L3域中富含腺的序列对于前Q1核糖开关功能至关重要.
研究的目的:
- 为了研究氨酸残留在preQ1 рибо开关的L3域中的作用.
- 评估2-aminopurine (2-AP) 替代对连接体结合和 рибо开关结构的影响.
- 了解L3的变化如何影响preQ1的识别.
主要方法:
- 对2-氨基氨酸 (2-AP) 的特定位点插入到前 Q1 核突变器的 L3 域.
- 光光谱学用于监测2-AP光.
- 循环二元化 (CD) 光谱分析全球形状变化.
- 联结体定位实验以确定结合亲和力.
主要成果:
- 在所有测试位置上,2-AP替代降低了preQ1结合亲和力.
- 全球形状变化发生在高度连接体的修饰和未修饰的 ribowitches 同样.
- 靠近连接体结合口袋的影响影响了2-AP对结合亲和力的影响.
- 远端2-AP位点在没有连接体的情况下影响了L3域的稳定性.
- 多次2-AP插入并没有造成进一步的重大破坏.
结论:
- 在L3域内的腺残留物对于识别preQ1联体是必不可少的.
- 2-AP替代提供了关于这些腺因残留物的复杂作用的见解.
- 这项研究突出显示了核突开关介导的基因调节中序列,结构和功能之间的复杂相互作用.
相关概念视频
01:56Riboswitches
8.1K
Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
8.1K
01:23Types of RNA
63.8K
Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
63.8K
02:24Ribosome Profiling
3.5K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.5K