Department of Physics, Indian Institute of Technology, Jammu, India.
Department of Physics, Indian Institute of Technology, Jammu, India; Department of Biosciences and Bioengineering, Indian Institute of Technology, Jammu, India.
Biophys J. 2024 Oct 15;123(20):3627-3639. doi: 10.1016/j.bpj.2024.09.014. Epub 2024 Sep 14.
Protein synthesis regulation primarily occurs at translation initiation, the first step of gene translation. However, the regulation of translation initiation under various conditions is not fully understood. Specifically, the reason why protein production from certain mRNAs remains resistant to stress while others do not show such resilience. Moreover, why is protein production enhanced from a few transcripts under stress conditions, whereas it is decreased in the majority of transcripts? We address them by developing a Monte Carlo simulation model of protein synthesis and ribosome scanning. We find that mRNAs with strong Kozak contexts exhibit minimal reduction in translation initiation rate under stress conditions. Moreover, these transcripts exhibit even greater resilience to stress when the scanning speed of 43S ribosome subunit is slow, albeit at the cost of reduced initiation rate. This implies a trade-off between initiation rate and the ability of mRNA to withstand stress. We also show that mRNAs featuring an upstream ORF can act as a regulatory switch. This switch elevates protein production from the main ORF under stress conditions; however, minimal to no proteins are produced under the normal condition. Because, in stress, a larger fraction of 43S ribosomes bypasses the upstream ORF due to its weak Kozak context. This, in turn, increases the number of scanning ribosomes reaching the main ORF, whose strong Kozak context can convert them into 80S ribosomes, even under stress conditions. This switching allows an efficient use of cellular resources by producing proteins when they are required. Thus, our computational study provides valuable insights into our understanding of stress-responsive translation-initiation.
蛋白质合成的调控主要发生在翻译起始阶段,即基因翻译的第一步。然而,在各种条件下,翻译起始的调控机制尚不完全清楚。具体来说,为什么某些 mRNA 产生的蛋白质对压力具有抗性,而其他 mRNA 则没有这种抗性;为什么在应激条件下,少数转录本的蛋白质产量增加,而大多数转录本的蛋白质产量减少。为了解决这些问题,我们开发了一种蛋白质合成和核糖体扫描的蒙特卡罗模拟模型。我们发现,具有强 Kozak 序列的 mRNA 在应激条件下,翻译起始速率的降低最小。此外,当 43S 核糖体亚基的扫描速度较慢时,这些转录本表现出更强的应激抗性,尽管这是以降低起始速率为代价的。这意味着在起始速率和 mRNA 承受应激的能力之间存在权衡。我们还表明,具有上游开放阅读框(ORF)的 mRNA 可以作为一种调节开关。这种开关在应激条件下提高了主 ORF 的蛋白质产量;然而,在正常条件下,几乎没有蛋白质产生。因为在应激条件下,由于其较弱的 Kozak 序列,较大比例的 43S 核糖体绕过上游 ORF。这反过来又增加了到达主 ORF 的扫描核糖体的数量,其强 Kozak 序列可以将它们转化为 80S 核糖体,即使在应激条件下也是如此。这种转换允许在需要时有效地利用细胞资源来产生蛋白质。因此,我们的计算研究为我们理解应激响应性翻译起始提供了有价值的见解。