ROLE OF THE P34CDC2 RELATED PITSLRE PROTEIN KINASES

P34CDC2 相关 PITSLRE 蛋白激酶的作用

• 批准号: 6194317
• 负责人: Mark Anthony Nelson
• 金额: $ 28.02万
• 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 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6194317
• 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（改编自研究者摘要）：复发性改变 1号染色体上的DNA是结构染色体上最常见的位点 所有人类实体瘤中的异常，包括人类皮肤 恶性黑素瘤在黑色素瘤中，涉及1号染色体的断裂点通常 累积到1 p36。编码PITSLRE蛋白激酶Cdc 2L 1的两个基因 和Cdc 2L 2定位于染色体带区1 p36。PITSLRE蛋白 激酶是p34 cdc 2超基因家族的一部分。若干条证据 表明PITSLRE蛋白激酶的某些亚型参与了 凋亡信号转导我们已经证明了Cdc 2L等位基因的缺失 黑色素瘤我们还可以证明PITSLRE的p110亚型被 半胱天冬酶转化为较小的p46同种型。p46同种型保留激酶活性， 似乎改变了它的底物特异性。也有区别 黑色素瘤细胞（具有野生型和突变型Cdc 2L等位基因）对 凋亡刺激。因此，要检验的主要假设是， 半胱天冬酶处理的PITSLRE同种型作为下游效应子在 细胞凋亡信号通路和PITSLRE蛋白激酶的破坏 在黑色素瘤的发展中起着功能性作用， 控制细胞死亡。三个具体目标是：（1）研究 Cdc 2L基因（PITSLRE）在黑色素瘤肿瘤进展过程中的改变 PCR-SSCP和DNA直接测序。档案散发性黑色素瘤病例 分析黑色素瘤的不同组织学阶段的特定突变， Cdc 2L 1基因中的区域。此外，家族性黑色素瘤激酶相关 到1 p也研究Cdc 2L 1基因的改变。(2)分离和 使用以下物质表征半胱天冬酶加工的PITSLRE同种型的底物： 酵母双杂交方法;（3）阐明 通过产生Cdc 2L 1缺陷小鼠使Cdc 2L基因失活。增加 了解引起人类免疫缺陷的关键基因改变， 黑色素瘤的发展和进展可能导致遗传标记， 黑素瘤定义Cdc 2L 1改变的重要性可用于识别 有趣的亚群Cdc 2L 1的遗传改变也可能具有预后意义。 与临床数据相结合时的价值。最后，增加 了解Cdc 2L的生物学作用可能会导致新的和创新的 治疗黑色素瘤的策略。