Targeting of proteins into peroxisomes

将蛋白质靶向过氧化物酶体

基本信息

批准号：
10612052
负责人：
JOHN D. AITCHISON
金额：
$ 68.88万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
1990
资助国家：
美国
起止时间：
1990-05-10 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10612052
关键词：
Address Alcohols Aldehydes Autophagocytosis Award Bile Acids Biogenesis Biological Process Biology Birth Cells Cellular biology Cholesterol Collaborations Communication Communities Comprehension Cues Development Diagnosis Disease Essential Genes Eukaryota Evolution Funding Future Genes Genetic Screening Glean Goals Grant Growth Homeostasis Human Hydrogen Peroxide Immunity Investigation Ions Laboratories Link Lipids Membrane Membrane Proteins Metabolic Pathway Mitochondria Molecular Biology Mutation Nature Nitrites Organelles Outcome Oxidation-Reduction Oxidative Phosphorylation Pathway interactions Patients Peroxisome Proliferation Plasmalogens Positioning Attribute Process Production Protein Import Proteins Quality Control Recording of previous events Research Rewards Role Signal Transduction Site Superoxides System Technology Time United States National Institutes of Health Vacuole Vesicle Viral Work Yeast Model System Yeasts disabling symptom experimental study fascinate high throughput screening innovation insight long chain fatty acid mutant novel oxidation peroxisome peroxisome membrane response success

项目摘要

We are excited, if presented with the opportunity, by the prospect of continuing over 3 decades of research into the mechanisms of peroxisome (PO) biogenesis. Our current work relies on yeast models, which have provided, and will continue to reveal, deep insights into this conserved and important biological process in humans, while also enhancing the diagnosis and understanding of the myriad of PO biogenesis disorders (PBDs). Our past work has centered on PO homeostasis, which balances the biogenesis and turnover processes, but we focus here on the birth of POs in cells that have no pre-existing POs, because previous genetic screens were done in cells that generate POs by redundant pathways involving both growth and division of pre-existing POs and de novo PO biogenesis. In doing so, redundant and essential genes were not identified, leaving a serious gap in our understanding. We remedy this using a novel, innovative, high-throughput screen (HTS) for PO biogenesis mutants in cells that are incapable of producing POs via growth and division. We show, using a pilot mini-screen, that unlike previous screens, our strategy is yielding putative hits in many steps of PO biogenesis, while identifying both known and novel genes, for the first time, including those involved in PO dynamics. This promising HTS platform, which we validate by proof-of-concept experiments, has provided a treasure trove of genes that now need deep investigation to understand how they function. Notably, many of the new genes we identified interact with known players we and others have been investigating, and a significant number are present at membrane contact sites (MCSs) between POs and other organelles. This presents a second fascinating avenue of pursuit, wherein we will probe the importance of metabolite (especially lipids) transport functions of the ER-PO MCSs. Our analyses will judiciously explore two yeast models to address evolutionary conservation and derive common, conserved principles. This leads naturally to the third, poorly-explored arena of interorganellar communication, for which we have found a novel, experimentally-tractable link, which is the requirement of mitochondrial redox and oxidative phosphorylation (OXPHOS) for PO proliferation (i.e. increase in PO number/cell). Thus, each of three new avenues of pursuit is expected to open new doors that will exceed what we alone can understand, but our long-term vision is to create interesting, new, community opportunities for future exploration in PO biology, knowing that it will be relevant, albeit indirectly, to patients with PBDs and the alleviation of their suffering. This proposal represents an expansion of scope by synergizing our molecular and cell biology efforts with the technical and intellectual prowess of the Aitchison laboratory (SCRI) and is backed by a significant track record of advances made by both PIs in PO biology. The Aims of our proposal are: Aim 1: Identify and perform functional studies on the machinery and regulators of de novo PO biogenesis. Aim 2: Elucidate how lipid regulators, MCSs and PO biogenesis proteins work together in ppV budding. Aim 3: Understand the role of mitochondrial signaling in the control of PO biogenesis.

如果有机会，我们会因继续研究超过3年的研究而感到兴奋过氧化物酶体（PO）生物发生的机制。我们目前的工作依赖于提供的酵母模型并将继续揭示人们对人类的保守和重要生物学过程的深刻见解，而还增强了对无数PO生物发生障碍（PBD）的诊断和理解。我们的过去工作以PO稳态为中心，该稳态平衡了生物发生和离职过程，但我们集中于关于POS在没有先前存在POS的细胞中的POS的诞生，因为先前的遗传筛选是在通过冗余途径产生POS的细胞，这些途径涉及POS和DE的生长和划分 Novo Po生物发生。这样做，尚未确定多余的基因，留下了严重的差距我们的理解。我们使用新颖的，创新的高通量屏幕（HTS）来解决此问题细胞中无法通过生长和分裂产生POS的突变体。我们显示，使用一个飞行员迷你屏幕，这与以前的屏幕不同，我们的策略在PO生物发生的许多步骤中产生了推定的命中，而首次识别已知基因和新型基因，包括参与PO动力学的基因。这我们通过概念验证实验来验证的有前途的HTS平台，提供了一个宝库现在需要深入研究以了解其功能的基因。值得注意的是，我们的许多新基因确定与已知玩家和其他人正在调查的已知玩家的互动，许多人是存在于POS和其他细胞器之间的膜接触位点（MCS）。这是一秒钟迷人的追求大道，其中我们将探索代谢物（尤其是脂质）运输的重要性 ER-PO MCSS的功能。我们的分析将明智地探索两个酵母模型以解决进化论保护和得出共同的保守原则。这自然会导致第三个，爆炸不良的竞技场我们找到了一个新颖的实验索环的组织间通信，这是线粒体氧化还原和氧化磷酸化（OXPHOS）的要求（即增加在PO数/单元格中。因此，预计三个新的追求途径将打开新的门，将超过我们一个人可以理解的是，我们的长期愿景是创造有趣的，新的，社区的机会对于PO生物学的未来探索，知道它与PBD的患者和减轻他们的痛苦。该提议通过协同分子来表示范围的扩展和细胞生物学努力，以Aitchison实验室（SCRI）的技术和知识能力为在PI在PO生物学中取得的重要进步的重大进展记录。我们提案的目的是：目标1：识别和执行有关从头生物发生的机械和调节剂的功能研究。目标2：阐明脂质调节剂，MCSS和PO生物发生蛋白如何在PPV萌芽中共同起作用。目标3：了解线粒体信号在控制PO生物发生中的作用。

项目成果

期刊论文数量（100）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Two independent pathways traffic the intraperoxisomal peroxin PpPex8p into peroxisomes: mechanism and evolutionary implications.

两条独立的途径将过氧化物酶体内过氧化物蛋白 PpPex8p 运输到过氧化物酶体中：机制和进化意义。

DOI：
10.1091/mbc.e05-08-0758
发表时间：
2006
期刊：
Molecular biology of the cell
影响因子：
3.3
作者：
Zhang,Lan;Léon,Sébastien;Subramani,Suresh
通讯作者：
Subramani,Suresh

Pex17p is required for import of both peroxisome membrane and lumenal proteins and interacts with Pex19p and the peroxisome targeting signal-receptor docking complex in Pichia pastoris.

Pex17p 是过氧化物酶体膜和腔蛋白输入所必需的，并与毕赤酵母中的 Pex19p 和过氧化物酶体靶向信号受体对接复合物相互作用。

DOI：
10.1091/mbc.10.12.4005
发表时间：
1999
期刊：
Molecular biology of the cell
影响因子：
3.3
作者：
Snyder,WB;Koller,A;Choy,AJ;Johnson,MA;Cregg,JM;Rangell,L;Keller,GA;Subramani,S
通讯作者：
Subramani,S

Phosphorylation-dependent Pex11p and Fis1p interaction regulates peroxisome division.