Alternative 18S rRNA 3' processing pathways in human ribosome biogenesis

人类核糖体生物合成中的替代 18S rRNA 3 加工途径

• 批准号: BB/R00143X/1
• 负责人: Claudia Schneider
• 金额: $ 45.11万
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FR00143X%2F1
• 关键词: Alternative; 18S; rRNA; processing; pathways

Life depends on the ability of an organism to react to changing environments and stresses by controlling the synthesis of complex macromolecules. Normal cell growth requires the production of ribosomes, the essential RNA-protein machinery that makes all cellular proteins. Ribosome assembly, a complicated process involving hundreds of biogenesis factors, is the major consumer of cellular energy and therefore tightly controlled. In cancer cells, ribosome production is upregulated to promote cell proliferation. Many cellular stresses and 20 genetic diseases ("ribosomopathies") result in a block or defect in ribosome production. Surprisingly, ribosomopathy patients are often pre-disposed to cancer and some cancers have also recently been linked to defective ribosome production. In humans, ribosome assembly controls the tumour suppressor p53, the "guardian of the genome". Through cellular signaling, p53 ensures appropriate cellular responses to stress conditions, and can even induce cell death if individual cells are detrimental to the whole organism. Indeed, p53 levels are elevated in many ribosomopathies and p53 is essential for many of the clinical features observed in these diseases.Here, we propose to study the impact of disease-associated mutations in key biogenesis factors on human ribosome production, p53 and cellular survival. Our internationally recognised expertise in ribosome biogenesis, RNA processing and cellular signaling, together with our strong preliminary data, mean that we are ideally placed to successfully carry out this project.Due to the high degree of evolutionary conservation in the ribosome production machinery, it was long assumed that ribosome production is the same in humans as it is in yeast, the main model organism used to study this process. However, we have now shown that the process of human ribosome production is significantly more complicated, and fundamentally different to that used in yeast. Surprisingly, human cells also retain the machinery needed for a "yeast-like" mechanism, however, this pathway is rarely used in humans and it is unclear what role it plays. Increasing our fundamental knowledge of human ribosome production is therefore of paramount importance for elucidating the basis of several human diseases and the development of potential treatments.WE WILL THEREFORE ADDRESS TWO KEY QUESTIONS:1) Do disease-associated mutations in key ribosomal RNA maturation factors cause defects or changes in ribosome production?2) Why do human cells use a specialised ribosomal RNA maturation pathway when they also have the machinery to employ a "yeast-like" mechanism?We will use a powerful combination of molecular tools and state-of-the-art technologies to reveal the mechanism(s) of human ribosome production and understand how this is altered in disease. This project will include determining the impact of disease-associated mutations on human ribosome production and p53-dependent cellular signaling, as well as using novel approaches to understand why human cells use a specialised ribosome production pathway. Intriguingly, our preliminary data also suggest that some cancer cells might depend on the minor, "yeast-like" mechanism. This raises the exciting possibility that this pathway could be targeted for the development of anti-cancer drugs in the future.

生命取决于生物体通过控制复杂大分子的合成来对不断变化的环境和压力做出反应的能力。正常的细胞生长需要核糖体的产生，核糖体是制造所有细胞蛋白质的基本RNA-蛋白质机器。核糖体组装是一个涉及数百个生物合成因子的复杂过程，是细胞能量的主要消耗者，因此受到严格控制。在癌细胞中，核糖体的产生被上调以促进细胞增殖。许多细胞应激和20种遗传性疾病（“核糖体病”）导致核糖体产生受阻或缺陷。令人惊讶的是，核糖体病患者通常易患癌症，最近一些癌症也与核糖体产生缺陷有关。在人类中，核糖体组装控制着肿瘤抑制基因p53，即“基因组的守护者”。通过细胞信号传导，p53确保对应激条件的适当细胞反应，并且如果单个细胞对整个生物体有害，甚至可以诱导细胞死亡。事实上，p53水平升高，在许多核糖体病和p53是必不可少的许多临床特征观察到在这些diseases.Here，我们建议研究疾病相关的突变的影响，在关键的生物合成因素对人类核糖体的生产，p53和细胞的生存。我们在核糖体生物合成、RNA加工和细胞信号传导方面的国际公认的专业知识，加上我们强大的初步数据，意味着我们是成功实施该项目的理想选择。由于核糖体生产机制的高度进化保守性，长期以来人们一直认为人类的核糖体生产与酵母中的核糖体生产相同，酵母是研究这一过程的主要模式生物。然而，我们现在已经证明，人类核糖体的生产过程要复杂得多，与酵母中使用的核糖体生产过程有根本的不同。令人惊讶的是，人类细胞也保留了“酵母样”机制所需的机制，然而，这种途径很少在人类中使用，目前还不清楚它起着什么作用。因此，增加我们对人类核糖体产生的基础知识对于阐明几种人类疾病的基础和开发潜在的治疗方法至关重要。我们将在此解决两个关键问题：1）关键核糖体RNA成熟因子中的疾病相关突变是否会导致核糖体产生的缺陷或变化？2)为什么人类细胞使用专门的核糖体RNA成熟途径时，他们也有机制，采用“酵母样”机制？我们将使用分子工具和最先进技术的强大组合来揭示人类核糖体产生的机制，并了解这在疾病中是如何改变的。该项目将包括确定疾病相关突变对人类核糖体产生和p53依赖性细胞信号传导的影响，以及使用新方法来理解为什么人类细胞使用专门的核糖体产生途径。有趣的是，我们的初步数据还表明，一些癌细胞可能依赖于次要的“酵母样”机制。这提出了一个令人兴奋的可能性，即这种途径可能成为未来抗癌药物开发的目标。

项目成果

RPS27a and RPL40, Which Are Produced as Ubiquitin Fusion Proteins, Are Not Essential for p53 Signalling.

• DOI: 10.3390/biom13060898
• 发表时间: 2023-05-28
• 期刊: Biomolecules
• 影响因子: 5.5

The induction of p53 correlates with defects in the production, but not the levels, of the small ribosomal subunit and stalled large ribosomal subunit biogenesis.

• DOI: 10.1093/nar/gkad637
• 发表时间: 2023-09-22
• 期刊: Nucleic acids research
• 影响因子: 14.9

Interactions and activities of factors involved in the late stages of human 18S rRNA maturation

人类 18S rRNA 成熟后期相关因素的相互作用和活动

• DOI: 10.6084/m9.figshare.7582253
• 发表时间: 2019
• 作者: Sloan K

RNA helicase-mediated regulation of snoRNP dynamics on pre-ribosomes and rRNA 2'-O-methylation.

• DOI: 10.1093/nar/gkab159
• 发表时间: 2021-04-19
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Aquino GRR;Krogh N;Hackert P;Martin R;Gallesio JD;van Nues RW;Schneider C;Watkins NJ;Nielsen H;Bohnsack KE;Bohnsack MT
• 通讯作者: Bohnsack MT

Nuclear stabilization of p53 requires a functional nucleolar surveillance pathway.

• DOI: 10.1016/j.celrep.2022.111571
• 发表时间: 2022-11-01
• 期刊: Cell reports
• 影响因子: 8.8