Defining a role for the mitochondrial protein sulfite oxidase in nucleolar ribosome biogenesis

定义线粒体蛋白亚硫酸氧化酶在核仁核糖体生物合成中的作用

基本信息

  • 批准号:
    10750182
  • 负责人:
  • 金额:
    $ 6.95万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2023
  • 资助国家:
    美国
  • 起止时间:
    2023-09-01 至 2025-08-31
  • 项目状态:
    未结题

项目摘要

PROJECT SUMMARY/ABSTRACT The mitochondrial protein sulfite oxidase (SUOX) has surprisingly emerged as a potential regulator of ribosome biogenesis in a genome-wide screen. Ribosome biogenesis, which occurs in the nucleolus of eukaryotic cells, is a highly regulated process essential for cell function. Despite its importance and relevance to diseases such as cancer and ribosomopathies, the regulation of ribosome biogenesis in human cells is not fully understood. To identify novel regulators of this process, the Baserga laboratory pioneered a genome-wide siRNA screen using the number of nucleoli per nucleus as an endpoint. MCF10A breast epithelial cells have an average nucleolar number of 2-3, and a decrease to one indicates aberrant ribosome biogenesis. SUOX is a mitochondrial protein that has unexpectedly surfaced as a hit from this screen. SUOX oxidizes toxic cellular sulfite to sulfate, and some SUOX variants are known to cause the severe, fatal developmental disease Isolated Sulfite Oxidase Deficiency (ISOD). Validation shows that SUOX depletion reduces nucleolar number and ribosomal RNA (rRNA) biogenesis. I have collected further data demonstrating ribosome biogenesis disruption upon SUOX depletion, and proteomics data strongly supporting a role for SUOX in making ribosomes. The similarity of the presentation of ISOD to that of known ribosomopathies, along with our intriguing preliminary results, has raised compelling questions about the involvement of ribosome biogenesis in ISOD pathogenesis. Drilling down to the mechanistic level, our preliminary metabolomics data reveal that SUOX depletion causes a decrease in the methyl donor required for the nucleolar methyltransferase fibrillarin. Fibrillarin methylates an rDNA-specific histone and rRNA, regulating rDNA transcription and rRNA processing, respectively. To date, no work has investigated the role of SUOX in ribosome biogenesis nor of ribosome biogenesis in ISOD pathogenesis. In Aim 1, I will establish the precise role of SUOX in ribosome biogenesis, taking advantage of established assays measuring multiple stages of the process. I will describe effects of SUOX depletion on rRNA methylation using the recently developed RibOxi-seq method, and histone methylation using established antibodies. I will further validate our results using the auxin-inducible degron version 2 (AID2) system. In Aim 2, I will determine the effects of disease-associated SUOX variants on human ribosome biogenesis by rescuing the defects that occur upon siRNA-mediated SUOX depletion with translationally silent and disease-associated loss-of-function SUOX variants. The experiments proposed will clearly define a role for SUOX in human ribosome biogenesis and describe the effects of known disease- causing variants on this essential cellular process. Our unique approach based on an unbiased screen for nucleolar function, combined with both well-established and novel methodology to study ribosome biogenesis, gives us the opportunity to take the field of SUOX research in an entirely new direction that will open avenues for understanding the function of SUOX in cellular metabolism and human genetic disease.
项目总结/摘要 线粒体蛋白亚硫酸盐氧化酶(SUOX)已令人惊讶地成为核糖体的潜在调节剂 在全基因组筛选中的生物发生。核糖体生物发生,发生在真核细胞的核仁中, 是细胞功能所必需的高度调节过程。尽管它的重要性和相关性的疾病, 与癌症和核糖体病一样,人细胞中核糖体生物发生的调节还没有完全了解。 为了确定这一过程的新调节因子,Baserga实验室率先进行了全基因组siRNA筛选 使用每个细胞核的核仁数作为终点。MCF 10A乳腺上皮细胞平均 核仁数为2-3,减少到1表明异常的核糖体生物发生。SUOX是一个 一种线粒体蛋白质,它意外地在这个屏幕上出现。SUOX氧化毒性细胞 已知某些SUOX变异体会导致严重的、致命的发育性疾病 孤立性亚硫酸氧化酶缺乏症(ISOD)。验证表明,SUOX耗竭减少核仁数 和核糖体RNA(rRNA)生物发生。我收集了更多的数据来证明核糖体的生物合成 蛋白质组学数据强烈支持SUOX在制造 核糖体ISOD与已知的核糖体病呈现的相似性,沿着我们的 有趣的初步结果,提出了令人信服的问题,核糖体生物合成参与 ISOD发病机制。深入到机制层面,我们的初步代谢组学数据显示, SUOX耗竭导致核仁甲基转移酶原纤蛋白所需的甲基供体减少。 Fibrillarin甲基化rDNA特异性组蛋白和rRNA,调节rDNA转录和rRNA加工, 分别到目前为止,还没有研究SUOX在核糖体生物合成中的作用,也没有研究核糖体的 ISOD发病机制中的生物发生。在目标1中,我将确定SUOX在核糖体生物发生中的确切作用, 利用已建立的测定方法测量该过程的多个阶段。我将描述 使用最近开发的RibOxi-seq方法对rRNA甲基化的SUOX消耗,以及组蛋白 使用已建立的抗体进行甲基化。我将进一步验证我们的结果使用生长素诱导降解决定子 版本2(AID 2)系统。在目标2中,我将确定疾病相关的SUOX变体对人类的影响。 通过用siRNA修复介导的SUOX耗尽后发生的缺陷来实现核糖体生物合成 功能性沉默和疾病相关的功能丧失SUOX变体。提出的实验将 明确定义SUOX在人类核糖体生物合成中的作用,并描述已知疾病的影响- 导致这一基本细胞过程的变异。我们独特的方法基于对 核仁功能,结合成熟的和新的方法来研究核糖体生物发生, 使我们有机会将SUOX研究领域带入一个全新的方向, 了解SUOX在细胞代谢和人类遗传疾病中的功能。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

Emily Catherine Sutton其他文献

Emily Catherine Sutton的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

相似海外基金

Modulation of Endogenous Auxins and Detoxification of Herbicidal Analogs in Plants by Gretchen Hagen 3 Enzymes
Gretchen Hagen 3 酶对植物内源生长素的调节和除草类似物的解毒
  • 批准号:
    558113-2021
  • 财政年份:
    2022
  • 资助金额:
    $ 6.95万
  • 项目类别:
    Postdoctoral Fellowships
Modulation of Endogenous Auxins and Detoxification of Herbicidal Analogs in Plants by Gretchen Hagen 3 Enzymes
Gretchen Hagen 3 酶对植物内源生长素的调节和除草类似物的解毒
  • 批准号:
    558113-2021
  • 财政年份:
    2021
  • 资助金额:
    $ 6.95万
  • 项目类别:
    Postdoctoral Fellowships
Study on the physiological roles of naturally occurring auxins in plants with distinct transport characteristics
具有独特运输特性的植物中天然存在的生长素的生理作用研究
  • 批准号:
    18H02457
  • 财政年份:
    2018
  • 资助金额:
    $ 6.95万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Next generation auxins and anti-auxins : principles for binding and design
下一代生长素和抗生长素:结合和设计原理
  • 批准号:
    BB/L010623/1
  • 财政年份:
    2014
  • 资助金额:
    $ 6.95万
  • 项目类别:
    Research Grant
Next generation auxins and anti-auxins : principles for binding and design
下一代生长素和抗生长素:结合和设计原理
  • 批准号:
    BB/L009366/1
  • 财政年份:
    2014
  • 资助金额:
    $ 6.95万
  • 项目类别:
    Research Grant
Distinct characteristics of two natural auxins in plants
植物中两种天然生长素的独特特征
  • 批准号:
    24370027
  • 财政年份:
    2012
  • 资助金额:
    $ 6.95万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Exploration of chemical probes for investigation of oxidative metabolism of auxins
生长素氧化代谢化学探针的探索
  • 批准号:
    22510228
  • 财政年份:
    2010
  • 资助金额:
    $ 6.95万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
RUI: Subcellular Basis of Plant Tissue Sensitivity to Auxins
RUI:植物组织对生长素敏感性的亚细胞基础
  • 批准号:
    9096239
  • 财政年份:
    1990
  • 资助金额:
    $ 6.95万
  • 项目类别:
    Standard Grant
RUI: Subcellular Basis of Plant Tissue Sensitivity to Auxins
RUI:植物组织对生长素敏感性的亚细胞基础
  • 批准号:
    9096327
  • 财政年份:
    1990
  • 资助金额:
    $ 6.95万
  • 项目类别:
    Standard Grant
Synthesis of Fungal potent auxins, accremoauxins, and their applied study on initiation and propagation of frail calli of conifers.
真菌强效生长素、accremoauxins的合成及其在针叶树脆弱愈伤组织引发和繁殖中的应用研究。
  • 批准号:
    02660128
  • 财政年份:
    1990
  • 资助金额:
    $ 6.95万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了