核心教条的单分子成像揭示了myosin-2A基因表达是由上下文翻译缓冲调节的
O'Neil Wiggan1, Timothy J Stasevich1,2
1Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO, 80525.
bioRxiv : the preprint server for biology
|February 19, 2024
概括
研究人员开发了一种新的细胞系来研究基因调节. 这使他们能够观察到MYH9基因表达如何在转录和翻译层面受到控制,揭示了称为翻译缓冲的机制.
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 基因规则 基因规则
背景情况:
- 蛋白质稳态对于细胞功能至关重要,并受到基因表达的严格调节.
- 传统的方法很难完全阐明维持蛋白质平衡的复杂调节机制.
- 编码髓-2A细胞骨架电机的MYH9基因是必不可少的,并且无处不在地表达.
研究的目的:
- 开发和利用一种新的CRISPR工程人类细胞系来剖析中央教条.
- 实时调查MYH9转录,翻译和蛋白质水平的动态.
- 揭示控制MYH9表达的调节机制,以响应细胞线索.
主要方法:
- 在内源MYH9基因中具有多个标签的人类细胞系的CRISPR工程.
- 同时对MYH9的转录,翻译和成熟的mRNA/蛋白质进行多色成像.
- 在不同细胞条件下的基因表达动态的量化,包括细胞骨干扰和血清刺激.
主要成果:
- MYH9转录通过细胞骨线索以SRF依赖的方式进行上调.
- MYH9的翻译效率可以动态调整以缓冲或匹配转录变化.
- 在科菲林被击倒后观察到翻译缓冲,减少了50%的翻译,以防止过度的髓活性.
- 翻译匹配了在血清刺激后的转录升高调节,以恢复稳定状态的MYH9水平.
结论:
- 语境翻译缓冲是确保稳定的MYH9表达的关键监管机制.
- 开发的细胞系提供了一个强大的工具来量化在各种细胞干扰下中央教条动态.
- 这种方法使我们能够全面了解基因调节和蛋白质平衡.
相关概念视频
01:23Regulation of Expression at Multiple Steps
907
The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
907
01:15Overview of Myosin Structure and Function
4.3K
Myosins are a family of molecular motor proteins, first identified in the skeletal muscles, where they are responsible for muscle contraction. Along with their role in muscle contraction, these proteins also play a role in the intracellular transport of molecules and vesicles. There are twenty-four classes of myosins based on their domain sequence and organization. Of the twenty-four, six classes (Myosin I, Myosin II, Myosin V, Myosin VI, Myosin VII, and Myosin X) have been well...
4.3K
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
02:24Regulation of Expression Occurs at Multiple Steps
22.7K
Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...
22.7K
02:22Regulated mRNA Transport
6.3K
In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing...
6.3K