Novel Functions of LAMMER-related Kinases in Gene Expression

Lammer相关蛋白在细胞生长、分化、乙肝病毒感染、 肿瘤发生与抗癌药物、胰岛素调节的葡萄糖代谢和营养的敏感性 动态平衡。蛋白水解酶家族的成员因其潜在的 作为各种人类疾病的治疗靶点。尽管它们的重要性和与疾病的相关性， 关于Lammer相关的激酶携带的机制，存在着相当大的知识空白 他们的职能。在先前关于它们在细胞周期控制和前mRNA剪接功能方面的工作的基础上， PI的实验室获得了初步数据，表明与Lammer相关的分裂酵母成员 Dsk1和Kic1/Lkh1在核输出和着丝粒基因沉默中的作用。的主要目标是 这项拟议的研究旨在破译Dsk1和Kic1激酶新角色背后的分子基础 在mRNA输出和着丝粒基因沉默中。这些实验旨在检验这样一个假设： Dsk1和Kic1可能通过调节Poly(A)结合活性在基因表达中发挥新的作用 蛋白质PABP参与着丝粒基因Swi6/HP1的mRNA输出和磷酸化级联 沉默。分子遗传学、生化和荧光显微镜将被用来评估活体 蛋白激酶对PABP和Swi6蛋白的磷酸化及细胞定位的影响 确定Dsk1/Kic1与PABP或Swi6可能的激酶-底物关系。裂变型酵母 庞氏裂殖酵母作为一个遗传和细胞生物学上易处理的小基因组系统 较少的冗余，为剖析多方面的激酶功能提供了一个有价值的模式生物。实现 这项研究的目标不仅将为深入了解Lammer激酶的分子机制提供帮助 在所有真核生物中发挥作用，但也有助于阐明所涉及的疾病的分子病理学和 解开更多开发治疗学的靶点。拟议的项目将为长期- 术语研究，以最终解决在多个步骤的激酶功能之间的相互作用 真核基因表达和细胞分裂周期，其保真度是癌症预防的中心问题 和治疗。此外，提案的教育目标与具体的 研究旨在将该项目融入基于课程的调查中，以增进学生对 项目主题，并鼓励妇女和科学界代表性不足的群体参与 研究。这个项目旨在为学生在这些领域的研究经验提供重要的机会。 与公众健康相关，激发他们对科学的兴趣，并使下一代专业人员多样化 在与健康相关的领域工作。拟议研究的意义和在 科学教育中的研究成分强化了凯克科学的研究环境 系和克莱蒙特学院。

LAMMER-related kinases play major roles in cell growth, differentiation, hepatitis B virus infection, cancer development and sensitivity to anti-cancer drugs, insulin-regulated glucose metabolism and nutrient homeostasis. Members of the kinase family are subjects of extensive interest because of their potential to serve as therapeutic targets for various human diseases. Despite their importance and disease relevance, considerable gaps of knowledge exist regarding the mechanisms by which the LAMMER-related kinases carry out their functions. Building upon previous work on their functions in cell cycle control and pre-mRNA splicing, the PI's laboratory obtained preliminary data that implicate the fission yeast members of the LAMMER-related kinases, Dsk1 and Kic1/Lkh1, in nuclear export of mRNA and centromeric gene silencing. The primary goal of the proposed research is to decipher the molecular basis underlying the novel roles of Dsk1 and Kic1 kinases in mRNA export and centromeric gene silencing. The experiments are designed to test the hypothesis that Dsk1 and Kic1 may execute their novel roles in gene expression by modulating the activity of poly(A)-binding protein Pabp in mRNA export and participating in Swi6/HP1 phosphorylation cascade for centromeric gene silencing. Molecular genetics, biochemical and fluorescence microscopy will be employed to assess in vivo phosphorylation and cellular localization of Pabp and Swi6 proteins affected by the kinases, as well as to determine the possible kinase-substrate relationship of Dsk1/Kic1 with Pabp or Swi6. The fission yeast Schizosaccharomyces pombe, as a genetically and a cell biologically tractable system with small genome size of less redundancy, offers a valuable model organism to dissect the multifaceted kinase functions. Achieving the goal of the research will not only provide insights into the molecular mechanisms of LAMMER kinase function in all eukaryotes but also contribute to elucidation of molecular pathology of the diseases involved and unraveling more targets for developing therapeutics. The proposed project will lay the foundation for the long- term investigation to ultimately address the interplay between the kinase functions in multiple steps of eukaryotic gene expression and cell-division cycle, the fidelity of which is the central issue of cancer prevention and treatment. Furthermore, the educational objectives of the proposal are intertwined with the specific research aims to integrate the project into course-based investigation, to enhance students' understanding of the project themes, and to encourage women and underrepresented groups in science to participate in research. This project aims to provide crucial opportunities for student research experience in the areas relevant to public health, stimulate their interest in sciences, and diversify the next-generation of professionals working in health-related fields. Both the significance of the proposed research and the enhancement in investigative components of science education strengthen the research environment of the Keck Science Department and the Claremont Colleges.