ROLES OF MSP58 IN NUCLEAR FUNCTION AND ONCOGENESIS

MSP58 在核功能和癌发生中的作用

• 批准号: 7715292
• 负责人: Hualin Zhong
• 金额: $ 11.89万
• 依托单位: HUNTER COLLEGE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7715292
• 关键词: Breast; Cause of Death; Cell Nucleus; Cell Proliferation; Cellular biology; Complex; Computer Retrieval of Information on Scientific Projects Database; Cultured Cells; Funding; Goals; Grant; Human; Immunoelectron Microscopy; Immunofluorescence Immunologic; Individual; Institution; Light; Location; Lung; Malignant Neoplasms; Malignant neoplasm of ovary; Maps; Mediating; Messenger RNA; Microscopy; Nuclear; Nuclear Export; Nuclear Pore Complex Proteins; Nucleolar Proteins; Oncogene Proteins; Pathogenesis; Play; Primary carcinoma of the liver cells; Process; Promoter Regions; Protein Export Pathway; Protein Overexpression; Protein Region; Proteins; Proto-Oncogenes; RNA Interference; Research; Research Personnel; Resources; Role; Role playing therapy; 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; fly; mRNA Differential Displays; mRNA Export; metaplastic cell transformation; novel; nucleocytoplasmic transport; 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. 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. 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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 癌症是美国和世界范围内的主要原因之一。许多癌症是由原癌基因的激活或抑癌基因的失活引起的。在大多数情况下，人们对这些癌蛋白/肿瘤抑制因子发挥作用的确切机制知之甚少。 Msp58（微球蛋白 58 kDa）是一种癌蛋白，从果蝇到人类在进化上都是保守的，并且已被证明在各种核过程中发挥作用，包括转录调节和 mRNA 输出。 Msp58 的过度表达已被证明可诱导细胞转化。 Msp58 执行其正常核功能的精确机制尚不清楚，也不了解为什么会异常表达 Msp58 导致细胞转化。该项目旨在阐明这两个问题。 我们已经鉴定出 Msp58 的两个新型相互作用伙伴：核孔蛋白 Tpr（易位启动子区域）和 E3 泛素连接酶 EDD（通过差异展示鉴定出 E3）。 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发挥重要作用。我们假设 Msp58 至少部分通过与 Tpr 相互作用来调节其一些关联因子在细胞核中的位置，从而发挥其功能。 EDD 通过其泛素直接或间接调节 Msp58 的稳定性和/或功能 连接酶活性。为了测试这些假设，在本项目中，我们将表征 Msp58 与 Tpr 和 EDD 的相互作用，并确定这些复合物的功能。删除分析和定点诱变将用于绘制介导相互作用的蛋白质区域。的地点 蛋白质和mRNA将通过免疫荧光共聚焦显微镜和免疫电子显微镜测定。我们将探索培养细胞中复合物各个成分的功能，其中成分的表达将被改变，要么通过 RNA 干扰沉默，要么过度表达 异位的。该项目的长期目标是探索Msp58的过度表达如何导致细胞转化。鉴于 Msp58 的相互作用蛋白，例如增殖相关核仁蛋白 p120 和 EDD，通常在各种癌症（包括肝细胞癌）中异常表达，并且 因此，这项研究不仅将增进我们对核组织和核运输基础细胞生物学的理解，而且有助于了解癌症的发病机制。