Investigating the role of HSC70 AMPylation on nuclear localization and chaperone function

研究 HSC70 AMPylation 对核定位和伴侣功能的作用

基本信息

批准号：
10231454
负责人：
Shannon Marie Lacy
金额：
$ 3.79万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
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：
2021-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10231454
关键词：
ATP phosphohydrolase Acute Adenosine Monophosphate Age Amyloid fibers Autophagocytosis Autophagosome Binding Biological Assay Caenorhabditis elegans Cardiac Cardiac Myocytes Cardiovascular Diseases Cause of Death Cell Culture Techniques Cell Fractionation Cell Line Cell Nucleus Cell physiology Cells Cellular Stress Cessation of life Coronary heart disease Cytosol Data Defect Defense Mechanisms Dysmyelopoietic Syndromes Failure Family Goals Heart Heart Diseases Heat Stress Disorders Heat shock proteins Heat-Shock Proteins 70 Heat-Shock Response Hypoxia Immunofluorescence Microscopy Impairment In Vitro Influenza Ischemia Knock-out Knockout Mice Knowledge Link Luciferases Mediating Modeling Molecular Molecular Chaperones Movement Mutate Myocardial Ischemia Nuclear Nuclear Import Nuclear Localization Signal Nuclear Protein Nuclear Translocation Orthologous Gene Patients Pharmacologic Substance Physiology Post-Translational Protein Processing Process Protein S Proteins Quality Control Recycling Regulation Risk Role Signal Transduction Site Stress System Testing Therapeutic Threonine Transgenic Organisms Work base cardiovascular risk factor chaperonin heart function misfolded protein novel nucleocytoplasmic transport prevent protein aggregation protein degradation protein folding proteostasis recruit response

项目摘要

Project Summary Coronary heart disease, also called Ischemic Heart Disease (IHD), is the leading cause of death worldwide. Repeated episodes of cardiac stress in IHD patients leads to an increase in misfolded proteins in their hearts. In response to this acute onset of cellular stress, heat shock proteins (HSP)s are upregulated in myocardiocytes and help restore protein homeostasis (proteostasis). Heat shock cognate 71 kDa (HSC70) is an essential, HSP70 chaperone that is located in the cytosol in the absence of cellular stress. This ATP-dependent chaperone actively refolds, degrades, and disaggregates proteins. HSC70 is also critical for autophagy, a cellular recycling/degradation process that protects the heart from misfolded proteins following ischemia. Impaired HSC70 function has been implicated in heart disease, yet despite the critical role of HSC70 in cellular physiology, the molecular mechanisms that regulate nuclear transport and govern the disparate functions of HSC70 remain unknown. Protein AMPylation recently emerged as a distinct PTM that regulates HSP70 family chaperones through the addition of adenosine monophosphate (AMP) to threonine residues. This process is catalyzed by the mammalian AMPylase, HYPE. My preliminary work revealed that HYPE AMPylates HSC70 on two sites, and increased cellular AMPylation levels prevent the nuclear-cytosolic shuttling of HSC70 as well as its ATPase activity. I also found that, similar to decreased levels of the HSC70 ortholog HSP-1, increased AMPylation inhibits autophagosome formation in C. elegans. These data suggest that HYPE-mediated AMPylation of HSC70 inhibits its critical functions associated with maintaining proteostasis, including nuclear-cytosolic shuttling, protein folding, and autophagy. The goals of this proposal are to 1) define the functional impact of HYPE-mediated AMPylation on HSC70 and 2) determine the impact of AMPylation on HSC70’s nuclear-cytosolic shuttling mechanism. Ultimately, we aim to define how a novel post-translational modification (PTM), AMPylation, regulates HSC70 function in proteostasis and cardiovascular diseases. Advancing our knowledge of the mechanisms that control HSC70 function is critical to identify novel avenues to therapeutic strategies that capitalize on restoring HSC70 regulation.

项目摘要冠心病，也称为缺血性心脏病（IHD），是全球死亡的主要原因。 IHD患者心脏应激的反复发作会导致他们心中折叠的蛋白质增加。在对这种细胞应激的急性发作的反应，热激蛋白（HSP）在心肌细胞中上调并有助于恢复蛋白质稳态（蛋白质稳定）。热冲击同源71 kDa（HSC70）是必不可少的在没有细胞应激的情况下，位于细胞质中的HSP70伴侣。这个依赖ATP的伴侣积极反映，降解和分解蛋白质。 HSC70对于自噬也很重要，一种细胞局部缺血后，可保护心脏免受错误折叠的蛋白质的回收/降解过程。受损 HSC70功能已在心脏病中隐含，但是尽管HSC70在细胞生理学中的重要作用，但调节核转运和控制HSC70不同功能的分子机制仍然存在未知。蛋白质氨基作最近作为一种独特的PTM出现，该PTM通过将腺苷一磷酸（AMP）添加到苏氨酸残留物中。这个过程是由哺乳动物催化的安培酶，炒作。我的初步工作表明，在两个地点上炒作HSC70，并增加细胞固定水平阻止了HSC70及其ATPase活性的核溶质穿梭。我也是发现类似于HSC70直系同源性HSP-1的水平降低，抑制了两臂。秀丽隐杆线虫中的自噬体形成。这些数据表明HSC70的炒作介导的两臂抑制它的关键功能与维持蛋白质量有关，包括核糖穿梭，蛋白质折叠和自噬。该提案的目标是1）定义炒作介导的两臂对HSC70和HSC70和 2）确定两极化对HSC70核溶质穿梭机制的影响。最终，我们的目标是定义新型翻译后修饰（PTM）如何调节蛋白质的HSC70功能和心血管疾病。促进我们对控制HSC70功能的机制的了解至关重要确定利用恢复HSC70法规的治疗策略的新途径。