The role of the NIMA kinase in mitotic regulation

NIMA 激酶在有丝分裂调节中的作用

• 批准号: 8206580
• 负责人: STEPHEN A OSMANI
• 金额: $ 37.74万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8206580
• 关键词: Affect; Affinity Chromatography; Animal Model; Aspergillus; Aspergillus nidulans; Biological Models; Biology; Cell Cycle; Cell Cycle Regulation; Cell Nucleolus; Cells; Chromatin; Complex; Congenital Abnormality; Coupled; DNA; DNA Binding; DNA Binding Domain; Data; Defect; Disease; Drosophila genus; Event; Genetic Transcription; Human; Importins; Integral Membrane Protein; Interphase; Kidney Diseases; Location; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Mediating; Medical; Medical Research; Medicine; Membrane Proteins; Messenger RNA; Mitosis; Mitotic; Modeling; Molds; NIMA; Nuclear Envelope; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Peripheral; Pharmaceutical Preparations; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Play; Positioning Attribute; Process; Proteins; Public Health; RNA Transport; Regulation; Research; Role; S Phase; Structure; Testing; Transmembrane Domain; Vertebrates; cancer therapy; cell killing; combat; in vivo; insight; mRNA Export; member; novel; pathogen; programs; public health relevance; research study; segregation; trafficking; tumor progression

DESCRIPTION (provided by applicant): Incorrect regulation of mitosis contributes to progression of cancer and birth defects and discoveries regarding the regulation of mitosis have positively impacted medical research and disease treatments. However, the potential for new insights and further impact on medicine clearly exists. For example, one of the most dramatic yet least understood mitotic processes is the disassembly and reassembly of nuclear pore complexes (NPCs), massive structures providing regulated gateways across the nuclear envelope. How this process is regulated and integrated with other mitotic events potentially involves reversible phosphorylation and our planned experiments will help test this hypothesis. We have shown the NIMA mitotic kinase of Aspergillus nidulans initiates mitosis by promoting NPC disassembly. NIMA is both required and sufficient to promote NPC disassembly and markedly affects mitosis in vertebrate cells suggesting the existence of conserved mitotic substrates. A. nidulans therefore provides a powerful and sophisticated model system in which to understand mitotic regulation of NPCs. The aims of this application include defining the dynamic mitotic changes in the NPC interactome using affinity purifications and mass spectrometry to identify new NPC proteins (Nups) as well as Nup associated proteins involved in mitotic regulation. In addition we will also focus studies on a new Nup associated protein (AN0162) which contains a transmembrane domain and potentially encodes a fourth transmembrane Nup. Our data suggest that AN0162 has a mitotic function to maintain the peripheral mRNA export factor Gle1 on the nuclear envelope surrounding the nucleolus during mitosis and we will test this hypothesis. Our third focus is on the mitotic functions of Nup2 and its associated proteins NupA, importin 1 and importin 2. Nup2 and NupA translocate from NPCs to chromatin during mitosis when they also become phosphorylated. We will test the hypothesis that NupA mediates the mitotic chromatin location of Nup2, which in turn controls the functions of importin 1 and 2 to regulate mitotic progression. As phosphorylation appears integral to this process, the final aim is to map mitotic phosphorylation sites within Nup2, NupA and importin 2 and then carry out in vivo mutational analysis of the specific phosphorylation sites to determine their function. PUBLIC HEALTH RELEVANCE: How the cell cycle is regulated is of fundamental medical importance because when this regulation goes awry disease states such as birth defects and cancer can result. In addition, many current cancer treatments target different aspects of cell cycle regulation to preferentially kill cells actively passing through the cell cycle. Therefore in terms of public health, the more we understand how the cell cycle is regulated the better positioned we will be to develop drugs against new chemotherapeutic targets to combat diseased states associated with cell cycle defects. The experiments proposed will in addition further our understanding of the NIMA kinase human orthologues of which are involved in specific disease states (kidney disease). Finally, A. nidulans is the model organism of the Aspergilli. Insights to the biology of A. nidulans will impact research on the opportunistic Aspergillus pathogens as well as members being used to generate new pharmacologically active compounds.

描述（由申请人提供）：有丝分裂的错误调节有助于癌症的进展，以及有关有丝分裂调节的出生缺陷和发现，对医学研究和疾病治疗产生了积极影响。但是，显然存在新见解和对医学的进一步影响的潜力。例如，最鲜为人知的有丝分裂过程之一是对核孔复合物（NPC）的拆卸和重新组装，巨大的结构可在整个核包膜上提供调节的网关。如何调节该过程并与其他有丝分裂事件进行整合，可能涉及可逆的磷酸化和我们计划的实验将有助于检验该假设。我们已经展示了曲霉尼古拉斯的NIMA有丝分裂激酶通过促进NPC拆卸来启动有丝分裂。 NIMA既需要又足以促进NPC拆卸，并显着影响脊椎动物细胞的有丝分裂，表明存在保守的有丝分裂底物。 A. nidulans因此提供了一个强大而复杂的模型系统，可以在其中了解NPC的有丝分裂调节。该应用程序的目的包括使用亲和力纯化和质谱法来定义NPC Intercontome中的动态有丝分裂变化，以鉴定新的NPC蛋白（NUP）以及参与有丝分裂调节的NUP相关蛋白。此外，我们还将重点研究新的NUP相关蛋白（AN0162），该蛋白包含跨膜结构域，并可能编码第四个跨膜NUP。我们的数据表明，AN0162具有有丝分裂功能，可以在有丝分裂过程中维持周围核仁核包膜上的外周mRNA输出因子GLE1，我们将检验该假设。我们的第三个重点是NUP2及其相关蛋白NUPA，Importin 1和Importin2。NUP2和NUPA在有丝质磷酸化期间从NPC转移到染色体的有丝分裂功能。我们将检验以下假设：NUPA介导NUP2的有丝分裂染色质位置，后者反过来控制Importin 1和2的功能以调节有丝分裂进程。由于磷酸化似乎是该过程不可或缺的，因此最终目的是绘制NUP2，NUPA和Importin 2中的有丝分裂磷酸化位点，然后对特定磷酸化位点进行体内突变分析，以确定其功能。 公共卫生相关性：细胞周期的调节如何具有基本的医学重要性，因为当该法规变为有因素疾病状态（例如先天缺陷和癌症）。此外，许多当前的癌症治疗靶向细胞周期调节的不同方面，以优先杀死细胞通过细胞周期。因此，就公共卫生而言，我们越了解如何调节细胞周期，我们将针对新的化学治疗靶标开发药物来打击与细胞周期缺陷相关的患病状态的药物。提出的实验还将进一步了解我们对特定疾病状态（肾脏疾病）的NIMA激酶人类直系同源物的理解。最后，A。Nidulans是曲霉的模型生物。对A. nidulans生物学的见解将影响对机会性曲霉病原体的研究，以及用于生成新的药理学活性化合物的成员。