ROLE OF THE P34CDC2 RELATED PITSLRE PROTEIN KINASES

P34CDC2 相关 PITSLRE 蛋白激酶的作用

• 批准号: 7232138
• 负责人: Mark Anthony Nelson
• 金额: $ 6.69万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-05-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7232138
• 关键词: apoptosis; cell cycle proteins; cysteine endopeptidases; enzyme activity; enzyme induction /repression; gene mutation; gene targeting; genetic markers; genetically modified animals; isozymes; laboratory mouse; melanocyte; melanoma; metastasis; neoplasm /cancer genetics; neoplastic process; nucleic acid sequence; polymerase chain reaction; protein kinase; single strand conformation polymorphism

Description (Adapted from the investigator's abstract): Recurring alterations of chromosome 1 represent the most frequent site of structural chromosome abnormalities across all human solid tumors, including human cutaneous malignant melanoma. In melanoma, breakpoints involving chromosome 1 often accumulate at 1p36. The two genes encoding the PITSLRE protein kinases, Cdc2L1 and Cdc2L2, are localized to chromosome band region 1p36. The PITSLRE protein kinases are part of the p34cdc2 supergene family. Several lines of evidence suggests that some of the PITSLRE protein kinases isoforms are involved in apoptotic signal transduction. We have demonstrated deletion of Cdc2L alleles in melanoma. We can also show that the p110 isoform of PITSLRE is cleaved by caspases to a smaller p46 isoform. The p46 isoform retains kinase activity but appears to change its substrate specificity. There is also a difference in the sensitivity of melanoma cells (with wildtype and mutant Cdc2L alleles) to apoptotic stimuli. Thus, the principal hypothesis to be tested is that the caspase-processed PITSLRE isoform functions as a downstream effector in apoptotic signaling pathways and that disruption of PITSLRE protein kinase function plays a functional role in melanoma development by deregulating controlled cell death. The three specific aims are: (1) To study the timing of Cdc2L gene (PITSLRE) alteration during melanoma tumor progression using PCR-SSCP and direct DNA sequencing. Archival sporadic melanoma cases from various histological stages of melanoma are analyzed for mutations of specific regions within the Cdc2L1 gene. In addition familial melanoma kindreds linked to 1p are also investigated for Cdc2L1 gene alterations. (2) To isolate and characterize the substrate for the caspase processed PITSLRE isoform using yeast two hybrid methodology and (3) To elucidate the biological function of Cdc2L gene inactivation by generating Cdc2L1 deficient mice. An increased understanding of the critical gene alterations that give rise to the development and progression of melanoma may lead to genetic markers for melanoma. Defining the importance of Cdc2L1 alterations may be used to identify interesting subpopulations. Genetic changes of Cdc2L1 also may have prognostic value when considered in tandem with clinical data. Finally, increased understanding of biological role of Cdc2L may lead to new and innovative strategies to treat melanoma.

描述（改编自调查员的摘要）：经常性更改 1染色体1是结构性染色体的最常见位点 所有人类实体瘤的异常，包括人皮肤 恶性黑色素瘤。在黑色素瘤中，经常涉及染色体1的断点 累积在136。编码Pitslre蛋白激酶Cdc2l1的两个基因 和cdc2l2局部局部到染色体带区域1p36。 Pitslre蛋白 激酶是p34CDC2超基因家族的一部分。几条证据 表明某些pitslre蛋白激酶同工型参与 凋亡信号转导。我们已经证明了CDC2L等位基因的删除 在黑色素瘤中。我们还可以证明pitslre的p110同工型被裂解 胱天蛋白酶达到较小的P46同工型。 P46同工型保留激酶活性，但 似乎改变了其底物特异性。在 黑色素瘤细胞（具有野生型和突变CDC2L等位基因）的敏感性 凋亡刺激。因此，要检验的主要假设是 caspase加工的Pitslre同工型在下游效应子中函数 凋亡信号通路和PITSLRE蛋白激酶的破坏 功能通过放松来在黑色素瘤发育中起作用 受控细胞死亡。三个具体目的是：（1）研究 黑色素瘤肿瘤进展过程中CDC2L基因（PITSLRE）的改变使用 PCR-SSCP和直接DNA测序。档案散发性黑色素瘤病例 分析了黑色素瘤的各种组织学阶段的特异性突变 Cdc2l1基因内的区域。另外，家族性黑色素瘤与之相关 还研究了1p的CDC2L1基因改变。 （2）分离和 表征使用使用使用的caspase加工叶子同工型的底物 酵母两种杂种方法和（3）阐明的生物学功能 通过产生Cdc2l1缺乏小鼠，Cdc2l基因失活。增加 理解引起的关键基因改变 黑色素瘤的发展和发展可能导致遗传标记 黑色素瘤。定义CDC2L1更改的重要性可用于识别 有趣的亚群。 cdc2l1的遗传变化也可能具有预后 与临床数据同时考虑时的价值。最后，增加了 了解CDC2L的生物学作用可能会导致新的创新性 治疗黑色素瘤的策略。