TULANE COBRE: PHOSPHORYLATION IN SOLID MUSCLE TUMOR ALVEOLAR RHABDOMYOSARCOMA

TULANE COBRE：实性肌肉肿瘤肺泡横纹肌肉瘤的磷酸化

• 批准号: 7382135
• 负责人: ANDREW DURRELL HOLLENBACH
• 金额: $ 29.49万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7382135
• 关键词: TULANE; COBRE; PHOSPHORYLATION; SOLID; MUSCLE

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. The solid muscle tumor Alveolar Rhabdomyosarcoma (ARMS), one of the most frequent soft tissue sarcomas in children, is characterized by the t(2;13)(q35;q14) chromosomal translocation, which results in the fusion of two transcription factors important for muscle development, Pax3 and FKHR. At present, little is known about the underlying molecular mechanisms regulating the activities of Pax3 and the oncogenic fusion protein Pax3-FKHR and how this regulation may be important for the development of ARMS. Therefore, the overall objective of this proposal is to understand how phosphorylation regulates the transcriptional activity of Pax3 and how Pax3-FKHR may alter this regulation. This objective will be addressed through two specific aims: (1) To investigate the role of phosphorylation in the regulation of Pax3 and Pax3-FKHR. This specific aim will be addressed by identifying the sites of phosphorylation on Pax3 and Pax3-FKHR, determining how phosphorylation affects the molecular biological activities of each protein (i.e.- DNA-binding, protein stability, and transcriptional activity), determining how phosphorylation alters the Pax3- and Pax3-FKHR-dependent enhanced proliferation and differentiation of primary myoblasts, and analyzing the importance of phosphorylation on the oncogenic activity of Pax3-FKHR. (2) To identify the kinases responsible for phosphorylating Pax3 and Pax3-FKHR. This specific aim wil''. be addressed by using standard chromatographic techniques to isolate the kinases that phosphorylate Pax3 and Pax3-FKHR from primary myoblasts, confirming that these kinases are responsible for the phosphorylation in vivo, and utilizing kinase inhibitors to determine how inhibition of these kinases effects the previously described Pax3 and Pax3-FKHR-dependent biological effects (i.e. - enhanced proliferation, enhanced apoptosis, and oncogenic activity). The comparison of the differences in the regulation of Pax3 and Pax3-FKHR by phosphorylation will provide important insights into the role of Pax3-FKHR in the formation of ARMS.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。实体肌肉肿瘤腺泡状横纹肌肉瘤 (ARMS) 是儿童中最常见的软组织肉瘤之一，其特征是 t(2;13)(q35;q14) 染色体易位，这导致两个转录因子的融合，这对于发育至关重要。肌肉发育、Pax3 和 FKHR。目前，人们对调节 Pax3 和致癌融合蛋白 Pax3-FKHR 活性的潜在分子机制以及这种调节对于 ARMS 的发展有何重要作用知之甚少。因此，本提案的总体目标是了解磷酸化如何调节 Pax3 的转录活性以及 Pax3-FKHR 如何改变这种调节。这一目标将通过两个具体目标来实现：(1) 研究磷酸化在 Pax3 和 Pax3-FKHR 调节中的作用。这一具体目标将通过识别 Pax3 和 Pax3-FKHR 上的磷酸化位点、确定磷酸化如何影响每种蛋白质的分子生物学活性（即 DNA 结合、蛋白质稳定性和转录活性）、确定磷酸化如何改变Pax3 和 Pax3-FKHR 依赖性增强原代成肌细胞的增殖和分化，并分析磷酸化对 Pax3-FKHR 致癌活性的重要性。 (2) 鉴定负责磷酸化Pax3和Pax3-FKHR的激酶。这个具体目标将会''。通过使用标准色谱技术从原代成肌细胞中分离磷酸化 Pax3 和 Pax3-FKHR 的激酶，确认这些激酶负责体内磷酸化，并利用激酶抑制剂来确定这些激酶的抑制如何影响先前描述的 Pax3 来解决该问题Pax3-FKHR 依赖性生物效应（即增强增殖、增强细胞凋亡和致癌活性）。比较 Pax3 和 Pax3-FKHR 磷酸化调节的差异将为了解 Pax3-FKHR 在 ARMS 形成中的作用提供重要见解。