CHARACTERIZATION OF MOB1 DYNAMICS IN LIVING CELLS

活细胞中 MOB1 动力学的表征

• 批准号: 7960229
• 负责人: Charles Bradley Shuster
• 金额: $ 12.87万
• 依托单位: NEW MEXICO STATE UNIVERSITY LAS CRUCES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7960229
• 关键词: Anaphase; Binding; Biomedical Research; CDC2 Protein Kinase; Cell division; Cells; Chimera organism; Chromosome Segregation; Chromosomes; Computer Retrieval of Information on Scientific Projects Database; Cytokinesis; Eukaryota; Family member; Foundations; Funding; Grant; Homologous Gene; Human; Institution; Kinetochores; LATS2 gene; Lead; Life; Maintenance; Mammalian Cell; Mitosis; Mitotic; Mitotic Checkpoint; Monitor; Names; New Mexico; Phosphotransferases; Play; Ploidies; Research; Research Personnel; Resources; Role; Signal Transduction; Sister Chromatid; Small Interfering RNA; Source; United States National Institutes of Health; Yeasts; cellular imaging; fungus; in vivo; mutant; novel; segregation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Maintenance of chromosomal ploidy during cell division requires a precise coordination of chromosome segregation and cytoplasmic partitioning (cytokinesis) such that the contractile ring does not assemble before the onset of anaphase. Progression of dividing cells into anaphase into cytokinesis is dependent on the spindle assembly checkpoint, which monitors chromosome attachment to the spindle and regulates the onset of sister chromatid segregation and CDK1 inactivation (mitotic exit). In yeast, the mitotic checkpoint also regulates a signaling cascade termed the Septation Initiation Network (SIN) that regulates the initiation of cytokinesis. Few functional homologues of the SIN have been identified in higher eukaryotes, but a terminal component named Mob1 has been identified in all eukaryotes. Expression of GFP chimeras of the four human Mob1s reveal that Mob1 localizes to the spindle poles and kinetochores up until anaphase onset, and the central spindle and midbody during cytokinesis. Additionally, we have identified a novel interaction between Mob1A and LATS2 kinase, which may serve as the functional homolog of the yeast Dbf2/Sid2 kinase in mammalian cells. Using these preliminary studies as a foundation, this application seeks to evaluate the roles that Mob1 plays in coordinating mitosis and cytokinesis in mammalian cells. The lines of experimentation proposed in this application will characterize the localization dynamics of the four human Mob1 family members using co-localization studies and live cell imaging. Further, the role of Mob1 in the coordination of mitosis and cytokinesis will be assessed by specifically disrupting Mob1/Lats2 interactions in vivo by siRNA knockdown of Mob1 or by expression of Lats2-binding mutants of Mob1. These efforts should ultimately lead to a clearer understanding of how mitosis and cytokinesis is coordinated, and confirm whether the strategies employed by unicellular fungi to regulate septation are conserved in higher eukaryotes.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 在细胞分裂过程中维持染色体倍性需要精确协调染色体分离和细胞质分配（细胞因子），以使收缩环在过后期开始之前不会组装。将细胞分为后期为细胞因子的进展取决于纺锤体组装检查点，该检查点可以监测染色体的附着在纺锤体上，并调节姐妹染色质分离和CDK1失活（有丝分裂出口）的发作。在酵母中，有丝分裂检查点还调节称为调节细胞因子启动的分离启动网络（SIN）的信号级联反应。在较高的真核生物中很少鉴定出罪的功能同源物，但是在所有真核生物中都鉴定了一个名为mob1的末端分量。四个人类MOB1的GFP嵌合体的表达表明，MOB1定位于主轴杆和动力学，直至原始原化，直到后期发作，以及胞质分裂期间的中央纺锤体和中体。此外，我们已经确定了MOB1A和LATS2激酶之间的新型相互作用，该相互作用可以作为哺乳动物细胞中酵母DBF2/SID2激酶的功能同源物。使用这些初步研究作为基础，该应用程序旨在评估MOB1在哺乳动物细胞中协调有丝分裂和细胞因子中的作用。本应用中提出的实验线将使用共定位研究和活细胞成像来表征四个人类MOB1家族成员的定位动力学。此外，将通过通过MOB1的siRNA敲低或通过表达MOB1的LATS2结合突变体的表达来评估MOB1在有丝分裂和细胞因子协调中的作用。这些努力最终应更清楚地了解有丝分裂和细胞因子如何协调，并确认单细胞真菌对调节分隔的策略是否在较高的真核生物中保存下来。