Identification of new regulators of centrosome positioning in C. elegans axis pol

描述(申请人提供)：在不同生物的发育过程中，作为分化过程的一部分，许多细胞变得极化。这一事件的一个例子发生在单细胞线虫胚胎中，这是一个建立前后(AP)体轴的过程。在许多类型的细胞中，这种极化事件是由中心体(细胞的微管组织中心)提示的。尽管许多工作涉及中心体，但人们对中心体在细胞中如何定位以确保极性建立知之甚少。PAM-1是线虫极性建立过程中中心体定位所必需的一种对嘌呤霉素敏感的氨基肽酶。嘌呤霉素敏感的氨基肽酶(PSA)广泛保守，在从植物到小鼠的生物体的减数分裂、细胞周期进展和繁殖中具有重要的发育作用。此外，PSA被认为在阿尔茨海默氏症和亨廷顿氏症等神经退行性疾病的发展中起到保护作用。这项拟议的工作旨在分离、表征和鉴定线虫中pam-1的抑制子。基因分析将被用来描述每个抑制子的性质。延时显微镜将被用来跟踪每个被抑制的菌株的中心体运动。单核苷酸多态图谱和全基因组测序将被用来识别每个抑制子突变。据推测，克隆和鉴定这些抑制突变将揭示与PAM-1协同工作的新蛋白，并在极性建立过程中作为氨基肽酶的靶标。鉴于PSA在神经退行性变中的意义，识别线虫中的靶标也可能促进对疾病机制的理解。此外，中心体定位的新参与者将被确定，这可能适用于其他系统。拟议的实验将主要由本科生进行，他们将得到指导，为他们提供在科学职业生涯和研究生工作中取得成功所需的指导和专业知识。

DESCRIPTION (provided by applicant): During development of diverse organisms, many cells become polarized as part of the differentiation process. One example of this event occurs in the one-cell C. elegans embryo, a process that establishes the anterior-posterior (AP) body axis. In many cell types, this polarization event is cued by the centrosome, the microtubule organizing center of the cell. Despite much work implicating the centrosome, little is known about how it is positioned in the cell to ensure polarity establishment. PAM-1 is a puromycin-sensitive aminopeptidase required for centrosome positioning during polarity establishment in C. elegans. Puromycin-sensitive aminopeptidases (PSA) are widely conserved and have important developmental roles in meiosis, cell cycle progression, and reproduction in organisms from plants to mice. Additionally, PSAs have been suggested to play a protective role in the development of neurodegenerative diseases such as Alzheimer's and Huntington's. The proposed work seeks to isolate, characterize and identify suppressors of pam-1 in C. elegans. Genetic analysis will be used to characterize the nature of each suppressor. Time-lapse microscopy will be used to track centrosome movements in each suppressed strain. Single-nucleotide polymorphism mapping and Whole genome sequencing will be employed to identify each suppressor mutation. It is hypothesized that cloning and characterization of these suppressor mutations will reveal novel proteins that work in conjunction with PAM-1 and as targets of the aminopeptidase during polarity establishment. Given the implication of PSAs in neurodegeneration, identification of targets in C. elegans may also advance the understanding of disease mechanisms. Additionally, new players in centrosome positioning will be identified which may be applicable to other systems. The proposed experiments will be largely carried out by undergraduates who will be mentored to provide them the guidance and expertise needed for success in science careers and graduate work.