Admin. supplement for equipment to Mechanisms underlying the Rlm1-dependent G1 checkpoint (NIH R15 GM135807)

行政。

基本信息

批准号：
10598250
负责人：
SAUL M HONIGBERG
金额：
$ 2.8万
依托单位：
UNIVERSITY OF MISSOURI KANSAS CITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10598250
关键词：
Biological Assay Bypass Cell Cycle Cell Cycle Progression Cell Cycle Regulation Cell Cycle Stage Cell Death Cell Maintenance Cell Wall Cell division Cell surface Cells Cellular Structures Chromosomes Cytology Disease Enzymes Equipment Event Genes Genetic High-Risk Cancer Lacazia loboi Lead Link Malignant Neoplasms Molecular Morphology Mutation Names Nature Pathogenesis Pathogenicity Positioning Attribute Proteins Role Signal Transduction Stress Structure Testing Time United States National Institutes of Health Work Yeasts anti-cancer therapeutic cancer cell cell injury daughter cell human pathogen mutant new therapeutic target novel parent grant programs repaired response transcription factor

Biological Assay Bypass Cell Cycle Cell Cycle Progression Cell Cycle Regulation Cell Cycle Stage Cell Death Cell Maintenance Cell Wall Cell division Cell surface Cells Cellular Structures Chromosomes Cytology Disease Enzymes Equipment Event Genes Genetic High-Risk Cancer Lacazia loboi Lead Link Malignant Neoplasms Molecular Morphology Mutation Names Nature Pathogenesis Pathogenicity Positioning Attribute Proteins Role Signal Transduction Stress Structure Testing Time United States National Institutes of Health Work Yeasts anti-cancer therapeutic cancer cell cell injury daughter cell human pathogen mutant new therapeutic target novel parent grant programs repaired response transcription factor

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项目摘要

(from parent grant) As cells divide the different cellular programs of cell division must be coordinated with one another. For example, the duplication of chromosomes, the separation of these duplicated chromosomes, and the division of one cell into two cells must all occur at precise times relative to one another and to other cellular events. To achieve this coordination, there are special times during the cell cycle, termed checkpoints, in which progression of the cell cycle can be paused. At these checkpoints, accurate/ undamaged completion of an earlier program is necessary before a later program will be initiated. Checkpoints work because the cell has specialized machines, made up of enzymes and other proteins, that are activated by damaged or incomplete cellular structures. Once activated these checkpoint machines cause the progression of the cell cycle to halt until the cellular structures are repaired and complete. When checkpoints are defective, cells continue through the cycle accumulating damaged structures and leading to disease states. Indeed, checkpoints are often bypassed in cancer cells, and defective checkpoints lead to higher risk of cancer. We recently proposed a novel checkpoint, here termed the “Rlm1-dependent checkpoint”, that delays passage through the cell cycle under a particular stressful environmental condition. As the name suggests, this checkpoint was revealed by a deletion mutant in the gene encoding the Rlm1 transcription factor. Rlm1 is known to be activated in response to cell-wall stress, so it may be that this checkpoint responds to stress at the cell surface. Interestingly, bypassing the Rlm1 checkpoint, unlike bypassing known checkpoints, leads to cell death in only one of the two products of cell division. Our long-term objective in this project is to characterize the function and mechanism of the Rlm1-dependent checkpoint. Our first specific aim is to characterize the position during the cell cycle at which this checkpoint operates, and the cell cycle regulators and additional cellular components through which it acts. Our second specific aim is to identify the nature of the signal to which this checkpoint responds and the nature of the damage that accumulates when the checkpoint is bypassed. To accomplish these aims we will employ a combination of genetic, cytological, and molecular biological assays. For example, we will grow both normal yeast and mutants defective in the Rlm1 checkpoint under non-stressful conditions and then release them suddenly into the stressful condition. We will then compare the normal and mutant yeast over time for known molecular or cellular events in the cell cycle as well as different types of cellular damage. We will test specific hypotheses regarding the mechanism of this checkpoint by examining mutants known to be defective in particular aspects of cell maintenance and cell cycle control for their role in this checkpoint. Characterization of this new type of checkpoint should reveal important new aspects of cell cycle control and may prove useful in understanding new aspects of disease states.

（来自父母赠款）随着细胞分裂，必须彼此协调细胞分裂的不同细胞程序。为了例如，染色体的重复，这些重复的染色体的分离和必须将一个细胞分成两个细胞的精确时间相对于彼此，而其他细胞则发生事件。为了实现这种协调，在细胞周期中有特殊时间，称为检查点，可以暂停细胞周期的哪个进展。在这些检查点，准确/未分娩在启动以后的程序之前，必须先进行较早的程序。检查点有效，因为细胞具有由酶和其他蛋白质组成的专用机器，这些机器被损坏或不完整的细胞结构。一旦激活这些检查点机，就会导致电池的进展循环停止，直到修复细胞结构并完成。当检查点有缺陷时，单元格继续整个周期，积累受损的结构并导致疾病状态。的确，检查点通常在癌细胞中绕过，并且检查点有缺陷导致癌症的风险更高。我们最近提出了一个新颖的检查站通过特定压力环境条件下的细胞周期。顾名思义，这个检查点由编码RLM1转录因子的基因中的缺失突变体揭示。 RLM1是已知会因细胞壁应力而被激活细胞表面。有趣的是，绕过RLM1检查点，与绕过已知检查点不同，线索仅在细胞分裂的两种产物之一中进行细胞死亡。我们在这个项目中的长期目标是表征RLM1依赖性检查点的功能和机制。我们的第一个具体目的是表征该检查点运行的单元格周期中的位置，并且细胞周期调节剂和其作用的其他细胞成分。我们的第二个具体目的是确定此检查点响应的信号的性质和累积损害的性质绕过检查点时。为了实现这些目标，我们将采用遗传的组合细胞学和分子生物学测定。例如，我们将同时生长正常的酵母和突变体在无压力条件下的RLM1检查点中有缺陷，然后突然释放到压力状况。然后，我们将随着时间的流逝比较正常和突变的酵母细胞周期中的细胞事件以及不同类型的细胞损伤。我们将测试特定通过检查已知在细胞维护和细胞周期控制的特定方面，以在此检查点中作用。这种新型检查点的表征应揭示细胞周期控制的重要新方面并可能证明有助于理解疾病状态的新方面。