The mammalian multi-tRNA synthetase complex
哺乳动物多tRNA合成酶复合物
基本信息
- 批准号:10531618
- 负责人:
- 金额:$ 48.19万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-12-01 至 2026-11-30
- 项目状态:未结题
- 来源:
- 关键词:3-DimensionalAllelesAmino AcidsAmino Acyl-tRNA SynthetasesArchitectureAtrophicBindingBiochemicalCell physiologyCellsCentral Nervous System DiseasesCerebrumChildComplexComputer softwareCoupledCritical ThinkingCrosslinkerCytoplasmCytoplasmic ProteinDNA Sequence AlterationDataData AnalysesDefectDiseaseEtiologyEventExhibitsFluorescenceFunctional disorderGenesGeneticGlutamine-tRNA ligaseGoalsGrantHealthHumanImpaired cognitionLengthLigaseMammalian CellMass Spectrum AnalysisMessenger RNAMethodsMicrocephalyModelingMolecularMultiprotein ComplexesMutateMutationMyelinNeurodegenerative DisordersNomenclaturePathologicPathologyPeptidesPhotobleachingPositioning AttributeProtein BiosynthesisProteinsReportingRibosomesRoleScaffolding ProteinSeizuresSodium ChlorideSpecificityStimulusStructural ModelsStructureSulfoxideTestingTranslationscrosslinkdensitydesignexperimental studyfluorescence imagingimprovedinsightinstrumentationinterestknock-downleukodystrophymRNA Translationmolecular modelingmotor impairmentneuropathologyprogramsprotein complexprotein crosslinksingle moleculespatial relationshipstoichiometrytool
项目摘要
Project Summary/Abstract
Mammalian cells contain a cytoplasmic multi-tRNA synthetase complex (MSC) consisting of 8 aminoacyl-tRNA
synthetases (AARSs) and 3 non-synthetase proteins. AARSs in the MSC function as “gene decoders” during
mRNA translation, but also exhibit non-canonical functions outside the MSC. However, the assembly, structure,
and function of the MSC are poorly understood. Importantly, mutations in genes encoding 7/11 constituents
cause central nervous system (CNS) disorders – five cause hypomyelinating leukodystrophy (HLD), and two
others cause progressive microcephaly. We will utilize state-of-the-art molecular approaches to improve our
understanding of the MSC, and its potential role in neuropathology. Our proposed Multiple-PI program takes
advantage of the expertise of two highly collaborative PI's – Paul Fox (Contact PI), a molecular biologist with
long-term interest in tRNA synthetases and the MSC, and Valentin Gogonea (Multiple PI), a physical chemist
with expertise in analysis and molecular modeling of multi-protein complexes. We will determine the quaternary
structure of the MSC by cross-linking mass spectrometry (XL-MS), a state-of-the-art method that facilitates
analysis of otherwise intractable complexes. To date we have found 19 inter-protein cross-links between all 11
MSC constituents, and 118 intra-protein cross-links. We have generated an initial model of the MSC that will
be refined here by XL-MS experiments with expanded amino acid specificity, and by SiMPull (single-molecule
pulldown) coupled with single-molecule fluorescence to determine stoichiometry. In addition, we will investigate
the mechanism of assembly of the MSC. Constitutive, multi-protein complexes are thought to be assembled by
domain-specific interactions between fully-formed, mature constituents (“post-translational assembly”).
However, assembly of some complexes utilizes a “co-translational assembly” mechanism in which a mature
constituent interacts with the nascent peptide of a partner constituent as it emerges from the ribosome. In
preliminary data we show at least 10 pairs of MSC constituents interact co-translationally. We will apply these
mechanistic approaches to elucidate the role of two MSC constituents in CNS diseases – genetic defects in
QARS1 and EPRS1 that cause microcephaly and HLD, respectively. Our preliminary studies indicate that
constituent mutation or suppression can lead to extra-MSC accumulation. Our preliminary studies have led us
to propose the following hypothesis: The mammalian MSC is a compact structure assembled in part by an
orderly sequence of co-translational interactions, however, mis-assembly or mutation can induce extra-MSC
accumulation of constituents, with potentially deleterious downstream consequences. We will test this
hypothesis by (1) determining MSC quaternary structure and component stoichiometry, and (2) determining the
role of co-translational interactions in MSC formation and integrity. We anticipate that elucidation of the
structure and assembly of the MSC will provide insights into mechanisms by which molecular defects in MSC
constituents can cause severe pathological disturbances, in particular, debilitating disorders of the CNS.
项目摘要/摘要
哺乳动物细胞含有一个细胞质多tRNA合成酶复合体(MSC),该复合体由8个氨基酰-tRNA组成
合成酶(AARs)和3种非合成酶蛋白。间充质干细胞中的AARs在
MRNA的翻译,但也表现出非典范功能外的MSC。然而,组装、结构、
对MSC的功能了解甚少。重要的是,编码7/11成分的基因突变
引起中枢神经系统(CNS)障碍--五种引起髓鞘减少性白质营养不良(HLD),两种
另一些则会导致进行性小头畸形。我们将利用最先进的分子方法来改善我们的
了解间充质干细胞及其在神经病理学中的潜在作用。我们建议的多PI计划需要
利用两位高度合作的派的专业知识-保罗·福克斯(联系派),一位具有
对tRNA合成酶和MSC的长期兴趣,以及物理化学家Valentin Gogonea(多PI)
在多蛋白质复合体的分析和分子建模方面具有专业知识。我们将确定第四纪
通过交联质谱(XL-MS)对MSC的结构进行分析,这是一种最先进的方法,有助于
对原本难以处理的复合体进行分析。到目前为止,我们已经在所有11个蛋白质之间发现了19个蛋白质间的交叉连接
MSC组分和118个蛋白质内交联物。我们已经生成了MSC的初始模型,它将
通过扩展氨基酸专一性的XL-MS实验和SiMPull(单分子)进行提纯
下拉)与单分子荧光联用以确定化学计量比。此外,我们还将调查
MSC的组装机制。构成的多蛋白质复合体被认为是由
完全形成的成熟成分之间特定领域的相互作用(“翻译后组装”)。
然而,一些复合体的组装利用了一种“共翻译组装”机制,在这种机制中,成熟的
当配对成分从核糖体中出现时,它与配对成分的新生多肽相互作用。在……里面
初步数据显示,至少有10对MSC成分相互作用。我们将应用这些
阐明两种间充质干细胞成分在中枢神经系统疾病中的作用的机制--遗传缺陷
QARS1和EPRS1分别引起小头畸形和肝豆状核变性。我们的初步研究表明
成分突变或抑制可导致MSC外积聚。我们的初步研究使我们
提出以下假设:哺乳动物的MSC是一个紧凑的结构,部分由一个
然而,有序的共翻译相互作用序列,错误组装或突变可以诱导额外的MSC
成分的积累,可能对下游造成有害后果。我们将对此进行测试
通过(1)确定MSC的四元结构和组分化学计量比,以及(2)确定
共翻译相互作用在MSC形成和完整性中的作用。我们期待对这一事件的澄清
MSC的结构和组装将提供对MSC中分子缺陷的机制的见解
成分可引起严重的病理障碍,特别是中枢神经系统的衰弱障碍。
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 }}
PAUL L FOX其他文献
PAUL L FOX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 }}
{{ truncateString('PAUL L FOX', 18)}}的其他基金
The Untranslated 3'End of SARS-CoV-2 RNA as a Determinant of Obesity-Accelerated Infectivity
SARS-CoV-2 RNA 的非翻译 3 末端是肥胖加速感染的决定因素
- 批准号:
10318871 - 财政年份:2021
- 资助金额:
$ 48.19万 - 项目类别:
The Untranslated 3'End of SARS-CoV-2 RNA as a Determinant of Obesity-Accelerated Infectivity
SARS-CoV-2 RNA 的非翻译 3 末端是肥胖加速感染的决定因素
- 批准号:
10689137 - 财政年份:2021
- 资助金额:
$ 48.19万 - 项目类别:
Assay Development for Discovery of a Small Molecule Inhibitor of a Novel Metabolic Pathway that Drives Obesity
发现导致肥胖的新型代谢途径的小分子抑制剂的检测方法开发
- 批准号:
10320035 - 财政年份:2020
- 资助金额:
$ 48.19万 - 项目类别:
Assay Development for Discovery of a Small Molecule Inhibitor of a Novel Metabolic Pathway that Drives Obesity
发现导致肥胖的新型代谢途径的小分子抑制剂的检测方法开发
- 批准号:
10115720 - 财政年份:2020
- 资助金额:
$ 48.19万 - 项目类别:
Multisite phosphorylated S6K1 directs a regulatory module determining adipocyte lipid metabolism
多位点磷酸化 S6K1 指导决定脂肪细胞脂质代谢的调节模块
- 批准号:
10349543 - 财政年份:2020
- 资助金额:
$ 48.19万 - 项目类别:
相似海外基金
Linkage of HIV amino acid variants to protective host alleles at CHD1L and HLA class I loci in an African population
非洲人群中 HIV 氨基酸变异与 CHD1L 和 HLA I 类基因座的保护性宿主等位基因的关联
- 批准号:
502556 - 财政年份:2024
- 资助金额:
$ 48.19万 - 项目类别:
Olfactory Epithelium Responses to Human APOE Alleles
嗅觉上皮对人类 APOE 等位基因的反应
- 批准号:
10659303 - 财政年份:2023
- 资助金额:
$ 48.19万 - 项目类别:
Deeply analyzing MHC class I-restricted peptide presentation mechanistics across alleles, pathways, and disease coupled with TCR discovery/characterization
深入分析跨等位基因、通路和疾病的 MHC I 类限制性肽呈递机制以及 TCR 发现/表征
- 批准号:
10674405 - 财政年份:2023
- 资助金额:
$ 48.19万 - 项目类别:
An off-the-shelf tumor cell vaccine with HLA-matching alleles for the personalized treatment of advanced solid tumors
具有 HLA 匹配等位基因的现成肿瘤细胞疫苗,用于晚期实体瘤的个性化治疗
- 批准号:
10758772 - 财政年份:2023
- 资助金额:
$ 48.19万 - 项目类别:
Identifying genetic variants that modify the effect size of ApoE alleles on late-onset Alzheimer's disease risk
识别改变 ApoE 等位基因对迟发性阿尔茨海默病风险影响大小的遗传变异
- 批准号:
10676499 - 财政年份:2023
- 资助金额:
$ 48.19万 - 项目类别:
New statistical approaches to mapping the functional impact of HLA alleles in multimodal complex disease datasets
绘制多模式复杂疾病数据集中 HLA 等位基因功能影响的新统计方法
- 批准号:
2748611 - 财政年份:2022
- 资助金额:
$ 48.19万 - 项目类别:
Studentship
Genome and epigenome editing of induced pluripotent stem cells for investigating osteoarthritis risk alleles
诱导多能干细胞的基因组和表观基因组编辑用于研究骨关节炎风险等位基因
- 批准号:
10532032 - 财政年份:2022
- 资助金额:
$ 48.19万 - 项目类别:
Recessive lethal alleles linked to seed abortion and their effect on fruit development in blueberries
与种子败育相关的隐性致死等位基因及其对蓝莓果实发育的影响
- 批准号:
22K05630 - 财政年份:2022
- 资助金额:
$ 48.19万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Investigating the Effect of APOE Alleles on Neuro-Immunity of Human Brain Borders in Normal Aging and Alzheimer's Disease Using Single-Cell Multi-Omics and In Vitro Organoids
使用单细胞多组学和体外类器官研究 APOE 等位基因对正常衰老和阿尔茨海默病中人脑边界神经免疫的影响
- 批准号:
10525070 - 财政年份:2022
- 资助金额:
$ 48.19万 - 项目类别:
Leveraging the Evolutionary History to Improve Identification of Trait-Associated Alleles and Risk Stratification Models in Native Hawaiians
利用进化历史来改进夏威夷原住民性状相关等位基因的识别和风险分层模型
- 批准号:
10689017 - 财政年份:2022
- 资助金额:
$ 48.19万 - 项目类别:














{{item.name}}会员




