Structural Basis for Translation Initiation in Leishmania Major
大利什曼原虫翻译起始的结构基础
基本信息
- 批准号:10373100
- 负责人:
- 金额:$ 25.4万
- 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:2021-03-16 至 2023-08-31
- 项目状态:已结题
- 来源:
- 关键词:AffinityAustriaBaculovirusesBindingBinding SitesBiochemicalBiochemistryBiological AssayBiophysicsBypassCellsCellular biologyClinicalComplexCountryCryoelectron MicroscopyCrystallizationCutaneousData CollectionDevelopmentDiseaseDorsalEIF4EL3 geneEffectivenessEndemic DiseasesEpitopesFaceFreezingFutureGene ExpressionGenesGoalsGuanosine TriphosphateHumanIndividualInfectionInterferometryLeishmaniaLeishmania majorLeishmaniasisLesionLife Cycle StagesMammalian CellMessenger RNAMethodsMini-ExonsMolecularMolecular ConformationMultiprotein ComplexesNMR SpectroscopyNucleic AcidsNucleotidesOutcomeParasite resistanceParasitesPathway interactionsPeptide Initiation FactorsPharmaceutical PreparationsPlayPreparationProcessProkaryotic Initiation Factor-3Protein BiosynthesisProtein IsoformsProtein Synthesis InhibitorsProteinsRNARNA BindingRNA Cap-Binding ProteinsRNA CapsRNA Recognition MotifReagentRecombinantsResolutionRibosomesRoentgen RaysRoleSamplingScaffolding ProteinScanningSeveritiesSideSiteSpliced Leader RNASpliced Leader SequencesStructureSurfaceSystemTetracyclinesTherapeutic InterventionTrans-SplicingTranscriptTranslation InitiationTranslationsTrypanosoma cruziVisceral LeishmaniasisWorkWorld Health Organizationanalogassay developmentbiophysical propertiesdrug developmentexperimental studyinducible gene expressioninhibitormedical schoolsneglected tropical diseasesprotein protein interactionrational designreconstitutionrecruitside effectsmall moleculesmall molecule inhibitorstructural biology
项目摘要
ABSTRACT
About 20 species of the protozoan parasite Leishmania are distributed across the globe and cause
roughly 1 million new cases of leishmaniasis annually. The most severe form of leishmaniasis (e.g., kala-azar or
visceral leishmaniasis) is lethal if untreated. Few therapies are available, and significant side effects and parasite
resistance limit their effectiveness. The World Health Organization lists Leishmaniasis as a neglected tropical
disease for which the development of new treatments is a high priority. Most of the gene expression in
Leishmania is regulated at the level of translation. In mammalian cells, translation initiation is well characterized.
A crucial step in this process includes the recognition by eIF4E, a cap-binding protein, of an m7GTP moiety
located at the 5’ end of messenger RNAs (mRNAs). eIF4E, through its interaction with other translation initiation
factors, ultimately coordinates the recruitment of the small ribosomal subunit.
To date, six Leishmania mRNA cap-binding protein isoforms (LIF4E-1 through -6) have been identified.
The Leishmania mRNA cap structure is unique and consists of the eukaryotic m7GTP cap followed by four
nucleotides that are hypermethylated (“cap-4”). A conserved mini-exon spliced leader RNA (SL RNA) of 39
nucleotides is also added to each transcript through trans-splicing. We previously determined the X-ray crystal
structure of LIF4E-1, the only cap-binding isoform that is expressed in Leishmania amastigotes (human infective
stage), bound to an interacting partner that represses its cap-binding activity (L4E-IP1). LIF4E-1 was also
recently shown to interact directly with the subunit “a” of Leishmania initiation factor 3 (LIF3), a large (~ 800 kDa)
multiprotein complex that binds the small ribosomal subunit. An interaction between an IF4E cap-binding protein
and an IF3 subunit has never been observed in other systems, and the details of the LIF4E-1/LIF3a molecular
interaction are unknown. We hypothesize that a LIF4E-1/cap-4 SL RNA/LIF3a interaction influences the
organization of LIF3 and how it assembles a competent pre-initiation complex (PIC) in amastigotes. The
LIF4E-1/LIF3a interaction would bypass the need for an eIF4G-like scaffolding protein. In this proposal, we will
pursue two specific aims: 1) To define the molecular basis for LIF3a interaction with the cap-binding protein
LIF4E-1. 2) To determine a high-resolution cryo-EM structure of LIF4E-1/cap-4 SL RNA bound to LIF3a or
assembled with LIF3 on the small ribosomal subunit. Our work will reveal the molecular basis for unique protein-
protein interactions in Leishmania parasites, and this information, in turn, could guide the development of specific
translation initiation inhibitors against these parasites.
摘要
大约20种原生动物寄生虫利什曼原虫分布在地球仪上,
每年大约有100万新的利什曼病病例。最严重的利什曼病形式(例如,黑热病
内脏利什曼病)如果不治疗是致命的。几乎没有可用的治疗方法,并且显著的副作用和寄生虫
阻力限制了其有效性。世界卫生组织将利什曼病列为被忽视的热带疾病
对于这种疾病,开发新的治疗方法是一个高度优先事项。大多数的基因表达在
利什曼原虫在翻译水平上受到管制。在哺乳动物细胞中,翻译起始被充分表征。
这个过程中的关键步骤包括eIF 4 E(一种帽结合蛋白)对m7 GTP部分的识别
位于信使RNA(mRNA)的5'末端。eIF 4 E,通过与其他翻译起始的相互作用
最终协调核糖体小亚基的募集。
迄今为止,已鉴定出六种利什曼原虫mRNA帽结合蛋白亚型(LIF 4 E-1至-6)。
利什曼原虫mRNA帽结构是独特的,由真核m7 GTP帽,随后是四个
高甲基化的核苷酸(“cap-4”)。一种39个保守的小外显子剪接前导RNA(SL RNA),
核苷酸也通过反式剪接添加到每个转录物中。我们之前确定了X射线晶体
LIF 4 E-1的结构,LIF 4 E-1是唯一在利什曼原虫无鞭毛体(人类感染性利什曼原虫)中表达的帽结合亚型。
阶段),与抑制其帽结合活性的相互作用伴侣(L4 E-IP 1)结合。LIF 4 E-1也是
最近显示直接与利什曼原虫起始因子3(LIF 3)的亚基“a”相互作用,LIF 3是一个大的(~ 800 kDa)
结合核糖体小亚基的多蛋白复合物。IF 4 E帽结合蛋白
在其他系统中从未观察到IF 3亚基,LIF 4 E-1/LIF 3a分子的细节
相互作用是未知。我们假设LIF 4 E-1/cap-4 SL RNA/LIF 3a相互作用影响了LIF 4 E-1/cap-4 SL RNA的表达。
LIF 3的组织以及它如何在无鞭毛体中组装一个有能力的前起始复合物(PIC)。的
LIF 4 E-1/LIF 3a相互作用将绕过对eIF 4G样支架蛋白的需要。在本提案中,我们将
本研究的目的有两个:1)确定LIF 3a与帽结合蛋白相互作用的分子基础
LIF4E-1 2)为了确定与LIF 3a结合的LIF 4 E-1/cap-4 SL RNA的高分辨率冷冻-EM结构,
在核糖体小亚基上与LIF 3组装。我们的工作将揭示独特蛋白质的分子基础-
利什曼原虫中的蛋白质相互作用,而这些信息反过来可以指导特异性
翻译起始抑制剂对这些寄生虫。
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 }}
Melissa Leger-Abraham其他文献
Melissa Leger-Abraham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('Melissa Leger-Abraham', 18)}}的其他基金
Ribosome structure determination from Apicomplexan parasites
顶复门寄生虫的核糖体结构测定
- 批准号:
10726704 - 财政年份:2023
- 资助金额:
$ 25.4万 - 项目类别:
Structural Basis for Translation Initiation in Leishmania Major
大利什曼原虫翻译起始的结构基础
- 批准号:
10225842 - 财政年份:2021
- 资助金额:
$ 25.4万 - 项目类别:
相似海外基金
Japanese Ethnogenesis: Oka Masao's Theory of Cultural Strata as a Hybrid Outcome of Scholarly Exchange between Austria and Japan
日本民族发生:冈正男的文化阶层理论是奥地利和日本之间学术交流的混合结果
- 批准号:
23K12262 - 财政年份:2023
- 资助金额:
$ 25.4万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
De-)constructing cultural colonialisms: A narratological investigation of Austria-Hungary's inner and extra-European discourses of identity and differ
解构文化殖民主义:对奥匈帝国内部和外部欧洲身份和差异话语的叙事学调查
- 批准号:
2877091 - 财政年份:2023
- 资助金额:
$ 25.4万 - 项目类别:
Studentship
The Research about the Non University Higher Education in Austria
奥地利非大学高等教育研究
- 批准号:
22K02686 - 财政年份:2022
- 资助金额:
$ 25.4万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
International Cooperative Research on Lesson Study-based Teacher Education between Japan, Germany, and Austria
日本、德国、奥地利课本式教师教育国际合作研究
- 批准号:
22KK0032 - 财政年份:2022
- 资助金额:
$ 25.4万 - 项目类别:
Fund for the Promotion of Joint International Research (Fostering Joint International Research (B))
IRES Track I: Exploring the Ecophysiology of Energy Balance in Vienna, Austria
IRES 第一轨:探索奥地利维也纳能量平衡的生态生理学
- 批准号:
1951995 - 财政年份:2022
- 资助金额:
$ 25.4万 - 项目类别:
Standard Grant
IRES Track II: Complexity advanced studies institute - Germany, Austria, Italy, Netherlands (Complexity-GAINs)
IRES Track II:复杂性高级研究机构 - 德国、奥地利、意大利、荷兰 (Complexity-GAINs)
- 批准号:
2106013 - 财政年份:2021
- 资助金额:
$ 25.4万 - 项目类别:
Standard Grant
How does the victimhood identity of right-wing populist parties in France, Germany, and Austria contribute to the rise in anti-Semitic hate crime?
法国、德国和奥地利右翼民粹主义政党的受害者身份如何导致反犹太仇恨犯罪的增加?
- 批准号:
2584860 - 财政年份:2021
- 资助金额:
$ 25.4万 - 项目类别:
Studentship
Europe's Last Peasant War: Violence and Revolution in Austria-Hungary and its Successors, 1917-1945
欧洲最后的农民战争:奥匈帝国及其继承者的暴力与革命,1917-1945 年
- 批准号:
AH/V004093/1 - 财政年份:2021
- 资助金额:
$ 25.4万 - 项目类别:
Research Grant
Implementation of incentive systems and nudges to promote environmentally conscious behavior ​and demonstration experiments in both Japan and Austria
实施激励制度和推动以促进环保意识行为
- 批准号:
21KK0027 - 财政年份:2021
- 资助金额:
$ 25.4万 - 项目类别:
Fund for the Promotion of Joint International Research (Fostering Joint International Research (B))
IRES Track 1: Neurobiology and evolution of frog dance displays in Austria and India
IRES 轨道 1:奥地利和印度青蛙舞蹈表演的神经生物学和进化
- 批准号:
1952542 - 财政年份:2020
- 资助金额:
$ 25.4万 - 项目类别:
Standard Grant