ROLES OF MSP58 IN NUCLEAR FUNCTION AND ONCOGENESIS

MSP58 在核功能和癌发生中的作用

• 批准号: 8357183
• 负责人: Hualin Zhong
• 金额: $ 12.28万
• 依托单位: HUNTER COLLEGE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357183
• 关键词: Basic Science; Cause of Death; Cell Differentiation process; Cell Nucleus; Cell Proliferation; Cellular biology; Center for Translational Science Activities; Complex; Confocal Microscopy; Cultured Cells; Funding; Goals; Grant; Human; Immunoelectron Microscopy; Immunofluorescence Immunologic; Individual; Light; Location; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of ovary; Maps; Mediating; Messenger RNA; National Center for Research Resources; Nuclear; Nuclear Export; Nuclear Pore Complex Proteins; Nucleolar Proteins; Oncogene Proteins; Pathogenesis; Play; Primary carcinoma of the liver cells; Principal Investigator; Process; Promoter Regions; Protein Region; Proteins; Proto-Oncogenes; RNA Interference; Research; Research Infrastructure; Resources; Role; Site-Directed Mutagenesis; Source; Testing; Transcriptional Regulation; Tumor Suppressor Proteins; Tumor-Suppressor Gene Inactivation; United States; United States National Institutes of Health; Work; cell transformation; cost; fly; health disparity; mRNA Differential Displays; malignant breast neoplasm; metaplastic cell transformation; novel; nucleocytoplasmic transport; overexpression; tumorigenesis; ubiquitin ligase; ubiquitin-protein ligase

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Cancer is one of the leading causes of death in the United States and in the world. Many cancers result from activation of proto-oncogenes or inactivation of tumor suppressor genes. In most cases, the precise mechanisms through which these oncoproteins/tumor suppressors work are poorly understood. Msp58 (microspherule protein 58 kDa), an oncoprotein, is evolutionarily conserved from fly to human and has been shown to play roles in various nuclear processes, including transcriptional regulation and mRNA export. Over-expression of Msp58 has been shown to induce cell transformation. The precise mechanism by which Msp58 carries out its normal nuclear functions is not known, nor is it understood why aberrant expression of Msp58 leads to cellular transformation. This project aims to shed light on both of these questions. We have identified two novel interacting partners of Msp58: nucleoporin Tpr (translocated promoter region) and E3 ubiquitin ligase EDD (E3 identified by differential display). Tpr has been implicated in nuclear organization and in the nuclear export of protein and messenger RNAs (mRNAs). EDD has been shown to play a critical role in cell proliferation and differentiation. We hypothesize that Msp58 carries out its functions by regulating the locations of some of its associating factors in the nucleus by, at least in part, interacting with Tpr; and EDD regulates the stability and/or the function of Msp58, directly or indirectly, through its ubiquitin ligase activity. To test these hypotheses, in this project, we will characterize the interaction of Msp58 with Tpr and EDD, and determine the functions of these complexes. Deletion analyses followed by site-directed mutagenesis will be used to map the regions of the proteins that mediate their interactions. The locations of proteins and mRNA will be determined by immunofluorescence confocal microscopy and immunoelectron microscopy. We will explore the functions of individual components of the complexes in cultured cells in which expression of the components will be altered, either silenced by RNA interference or overexpressed ectopically. A long-term goal of this project is to explore how Msp58's over-expression leads to cellular transformation. Given that Msp58's interacting proteins, such as proliferation-associated nucleolar protein p120 and EDD, are often aberrantly expressed in various cancers, including hepatocellular carcinoma, and breast, lung and ovarian cancers, this study, therefore, will not only advance our understanding of the fundamental cell biology underlying nuclear organization and nuclear transport, but also shed light on understanding the pathogenesis of cancers.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 癌症是美国和世界上主要的死亡原因之一。许多癌症是由原癌基因激活或肿瘤抑制基因失活引起的。在大多数情况下，这些癌蛋白/肿瘤抑制因子的确切机制尚不清楚。Msp 58（microspherule protein 58 kDa）是一种从果蝇到人类进化上保守的癌蛋白，在转录调控和mRNA输出等多种核过程中发挥重要作用。 已经显示Msp 58的过表达诱导细胞转化。Msp 58执行其正常核功能的确切机制尚不清楚，也不理解为什么Msp 58的异常表达会导致细胞核内的细胞凋亡。 Msp 58导致细胞转化。该项目旨在阐明这两个问题。 我们已经确定了两个新的相互作用的合作伙伴Msp 58：核孔蛋白Tpr（易位启动子区）和E3泛素连接酶EDD（E3确定的差异显示）。TPR与核组织以及蛋白质和信使RNA（mRNA）的核输出有关。EDD已被证明在细胞增殖和分化中起关键作用。我们推测Msp 58通过调节其相关因子在细胞核中的位置来实现其功能，至少部分通过与Tpr相互作用来实现; EDD通过其泛素直接或间接地调节Msp 58的稳定性和/或功能 连接酶活性为了验证这些假设，在本项目中，我们将表征Msp 58与Tpr和EDD的相互作用，并确定这些复合物的功能。将使用缺失分析，然后进行定点诱变，以绘制介导其相互作用的蛋白质区域。的位置 通过免疫荧光共聚焦显微镜和免疫电子显微镜测定蛋白质和mRNA。我们将探索培养细胞中复合物单个组分的功能，其中组分的表达将被改变，要么通过RNA干扰沉默，要么过表达 异位的该项目的长期目标是探索Msp 58的过度表达如何导致细胞转化。鉴于Msp 58的相互作用蛋白，如增殖相关核仁蛋白p120和EDD，经常在各种癌症中异常表达，包括肝细胞癌， 乳腺癌，肺癌和卵巢癌，因此，这项研究不仅将促进我们对核组织和核运输的基本细胞生物学的理解，而且还有助于了解癌症的发病机制。