Role of LIN-9 in Mammalian Cell Cycle Regulation

LIN-9 在哺乳动物细胞周期调节中的作用

• 批准号: 7907918
• 负责人: RAUDEL SANDOVAL
• 金额: $ 13.49万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-26 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7907918
• 关键词: Address; Amino Acids; Biochemical; CDK4 gene; Caenorhabditis elegans; Cell Cycle; Cell Cycle Proteins; Cell Cycle Regulation; Cell Line; Cell Proliferation; Cells; Clinical; Complex; Cyclin D1; Embryo; Fibroblasts; Funding; Genetic; Intention; Knockout Mice; Mammalian Cell; Maps; Mentors; Modification; Mus; Nature; Normal Cell; Orthologous Gene; Pathway interactions; Phase; Plasmids; Postdoctoral Fellow; Protein Family; Proteins; Recombinant Proteins; Regulation; Research Personnel; Role; Signal Transduction; Time; Translating; Work; anticancer research; cell growth; knowledge base; member; mutant; novel; novel therapeutic intervention; protein complex; tumor

DESCRIPTION (provided by applicant): At the present time, our work focuses on cell cycle regulation by a mammalian form of C. elegans lin-9. In C. elegans, lin-9 functions downstream of the mammalian equivalent of CDK4 in a pathway that regulates cell proliferation. We demonstrate that this lin-9 ortholog, referred throughout the proposal as BARA/LIN-9, also appears to act downstream of cyclin D/CDK4 in mammalian cells. This is supported by the fact that over- expression of BARA/LIN-9 has an inhibitory effect on cell proliferation which is partially blocked by co- expression of cyclin D1 and, furthermore, the partial rescue of phenotypic alterations in CDK-null mice by expression of a mutant form of BARA/LIN-9 lacking amino acids 1-84 (BARA/LIN-9?84). Recently, we have determined that BARA/LIN-9 interacts with p107/p130, members of the pocket protein family, and E2F4. These preliminary results are novel and extremely important in further illuminating our understanding of cell cycle regulation. Obviously, it is through alterations, typically through genetic modifications, of normal cell cycle function that leads to an unchecked state of cell growth, which ultimately promotes tumor formation. It is my intention to pursue this study further as I begin my transition from mentored postdoctoral fellow to independent investigator. To that end, this proposal, if funded, would allow me to pursue a more detailed study of the protein interactions between BARA/LIN-9, p107/p130 and E2F4 through studies that will involve protein mapping with recombinant proteins. Furthermore, I will study the stability of this complex temporally along the cell cycle and examine the potential interaction with other cell cycle proteins. Changes of this complex along different phases of the cell cycle will help us to better understand the potential signaling mechanisms that may regulate this protein complex. In addition, I will also use primary mouse fibroblasts (MEFs) from mice expressing BARA/LIN-9?84 to determine if the interactions with p107/p130 and E2F4 are altered. Moreover, I will continue to study the knockout (KO) mice we have generated to elucidate a mechanism for cyclin D/CDK4 regulation of LIN-9 function. Taken together, this study will produce a more enhanced understanding of cell cycle regulation, which will add substantially to our continuously growing knowledge base and aid the entire cancer research field as we continuously look for novel signaling mechanisms that may translate into new therapeutic approaches in the clinical field.

描述（由申请人提供）：目前，我们的工作集中于通过哺乳动物形式的秀丽隐杆线虫lin-9形式的细胞周期调节。在秀丽隐杆线虫中，LIN-9在调节细胞增殖的途径中，哺乳动物等效的哺乳动物等效的LIN-9功能。我们证明，在整个提案中称为bara/lin-9的LIN-9直系同源物也似乎在哺乳动物细胞中的细胞周期蛋白D/CDK4下游作用。这一事实支持了这一事实，即Bara/Lin-9的过度表达对细胞增殖具有抑制作用，而细胞增殖受到了Cyclin D1的共同表达而部分阻塞，此外，CDK-NULL小鼠中表型变化的部分挽救通过Bara/lin-9缺乏氨基酸的突变体表达的表型变化，而Saveling amino-9缺乏氨基氨基酸1-84（Bara Amino Amino Acion 1-84？84）？最近，我们确定Bara/Lin-9与Pocket蛋白家族和E2F4的P107/P130相互作用。这些初步结果是新颖的，对于进一步阐明我们对细胞周期调节的理解极为重要。显然，通常通过改变正常细胞周期功能的遗传修饰的改变，导致细胞生长的未检查状态，最终促进肿瘤的形成。当我开始从指导的博士后研究员到独立调查员的过渡时，我打算进一步进行这项研究。为此，该提案（如果资助）将使我可以通过研究将涉及与重组蛋白的蛋白质映射的研究进行更详细的研究，以对BARA/LIN-9，P107/P130和E2F4之间的蛋白质相互作用进行更详细的研究。此外，我将研究该复合物沿细胞周期的稳定性，并检查与其他细胞周期蛋白的潜在相互作用。该复合物沿细胞周期的不同阶段的变化将有助于我们更好地了解可能调节该蛋白质复合物的潜在信号传导机制。此外，我还将使用表达bara/lin-9？84的小鼠的主要小鼠成纤维细胞（MEF）来确定是否改变了与P107/P130和E2F4的相互作用。此外，我将继续研究我们已经生成的敲除（KO）小鼠，以阐明lin-9功能的细胞周期蛋白D/CDK4调节机制。综上所述，这项研究将产生对细胞周期调节的更高理解，这将大大增加我们不断增长的知识基础，并帮助整个癌症研究领域，因为我们不断寻找可以转化为临床领域的新治疗方法的新型信号机制。