Growth Arrest in Differentiating Keratinocytes

分化角质形成细胞的生长停滞

• 批准号: 6747852
• 负责人: DEEPAK K AGRAWAL
• 金额: $ 22.84万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6747852
• 关键词: cell differentiation; cell growth regulation; chimeric proteins; complementary DNA; cyclin dependent kinase; cyclins; enzyme activity; gene induction /repression; genetic promoter element; keratinocyte; laboratory mouse; luciferin monooxygenase; p53 gene /protein; protein protein interaction; recombinant proteins

DESCRIPTION (provided by applicant): It is well established that much of the complex regulation exerted during G1 and S phase traverse in eukaryotic cells is under the control of a family of latent protein kinases, cdks, which are activated by synthesis and binding of their positively acting cyclin subunits. By contrast, substantially less is known about the potential function of cyclin/cdks during terminal differentiation in mammalian cells. Like proliferation, differentiation occurs through a series of tightly controlled temporally regulated sequence of events. However, it is unclear if the same cyclin/cdks that have become an established paradigm in cell growth are the same that are responsible for regulating differentiation We have identified cyclin G2 as a strongly induced gene in murine keratinocytes forced to undergo differentiation in a suspension culture model and in suprabasal keratinocytes harvested from skin. This application seeks to identify the role of this cyclin in enforcing a growth arrest in murine keratinocytes during terminal differentiation. In the first specific aim, the proposed studies will seek to establish whether cyclin G2 functions in collaboration with the cyclin/cdks that exert replicative control in growing cells. These studies will utilize ectopic expression of G1 and S phase cyclins as well as ectopic cyclin G2 expression to ascertain if there is a functional relationship or if cyclin G2 functions through a parallel independent pathway. The studies described in the second specific aim will focus on identifying the factor(s) responsible for the transcriptional induction of cyclin G2 that we have discovered in keratinocytes. Additional investigations will determine the role of the p53 homolog p63 in cyclin G2 induction. The final specific aim will be directed at identifying cyclin G2 interacting proteins in order to identify a catalytic partner(s) for cyclin G2. These studies will be accomplished by using biochemical methods to isolate proteins with affinity for recombinant cyclin G2 and also by screening mammalian cDNA libraries using a two hybrid system.

描述（由申请人提供）： 众所周知，大部分 真核细胞 G1 和 S 期穿越过程中的复杂调控 受潜在蛋白激酶家族 cdks 的控制，该家族是 通过其积极作用的细胞周期蛋白亚基的合成和结合而被激活。 相比之下，人们对潜在功能知之甚少。 哺乳动物细胞终末分化期间的细胞周期蛋白/cdks。喜欢 增殖、分化是通过一系列严格控制的过程发生的 事件的时间顺序。但尚不清楚是否相同 细胞周期蛋白/cdks 已成为细胞生长的既定范例是 我们已经确定了负责调节分化的相同 细胞周期蛋白 G2 作为小鼠角质形成细胞中的强诱导基因，被迫经历 悬浮培养模型和基底上角质形成细胞中的分化 从皮肤中收获。该应用旨在确定该细胞周期蛋白的作用 在终末期强制小鼠角质形成细胞生长停滞 差异化。 在第一个具体目标中，拟议的研究将寻求确定是否 cyclin G2 与 cyclin/cdks 协同发挥作用，发挥作用 生长细胞中的复制控制。这些研究将利用异位 G1 和 S 期细胞周期蛋白的表达以及异位细胞周期蛋白 G2 的表达 确定是否存在功能关系或细胞周期蛋白 G2 是否发挥作用 通过平行的独立路径。第二部分中描述的研究 具体目标将侧重于确定造成问题的因素 我们发现细胞周期蛋白 G2 的转录诱导 角质形成细胞。进一步的研究将确定 p53 的作用 细胞周期蛋白 G2 诱导中的同源物 p63。最终的具体目标将针对 鉴定细胞周期蛋白 G2 相互作用蛋白以鉴定催化 细胞周期蛋白 G2 的伙伴。这些研究将通过使用来完成 生化方法分离对重组细胞周期蛋白 G2 具有亲和力的蛋白质 还通过使用两种杂交系统筛选哺乳动物 cDNA 文库。