CHARACTERIZATION OF AHR COMPLEX IN MALIGNANT TUMOR CELLS

恶性肿瘤细胞中 AHR 复合物的表征

• 批准号: 8365505
• 负责人: David H Sherr
• 金额: $ 0.46万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-08-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365505
• 关键词: ARNT gene; Affect; Aromatic Polycyclic Hydrocarbons; Aryl Hydrocarbon Receptor; Binding; Binding Sites; Biology; Cell Line; Cell Nucleus; Cells; Circadian Rhythms; Complex; DNA Binding; DNA-Binding Proteins; Data Analyses; Databases; Development; Dimerization; Dioxins; Family; Fetal Kidney; Funding; Gene Targeting; Genetic Transcription; Grant; Human; Ligand Binding; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Medicine; N-terminal; National Center for Research Resources; Neurologic; Normal Cell; Nuclear; Peptides; Principal Investigator; Process; Promoter Regions; Proteins; Research; Research Infrastructure; Resources; Sampling; Site; Source; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; United States National Institutes of Health; angiogenesis; bHLH Domain; cost; kidney cell; member; neoplastic cell; receptor; transcription factor

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. Most of the activity of environmentally ubiquitous polycyclic aromatic hydrocarbons (PAH) and related dioxin are mediated by the AhR (aryl hydrocarbon receptor). The AhR is a member of the Per/ARNT/Sim (PAS) family of transcription factors and is known to be associated with neurological development, circadian cycle and angiogenesis. The AhR contains a DNA binding and protein dimerization site in its N-terminal bHLH domain, a ligand binding site in the PAS homology domain, nuclear localization and export sequences, and a transcriptionally active Q-rich domain. Upon ligand binding, the receptor translocates into the nucleus and associates with at least one co-factor, ARNT. The activated AhR complex then binds promoter regions of a number of target genes at specific DNA-binding sites, resulting in gene transcription. If the AhR is constitutively active in tumor cells and its transcriptional activity affects functions which are critical to those cells, then it wou ld b e expected that the composition of the active AhR complex would be different in tumor cells than in their non-transformed counterparts. The purpose of this study is to identify and compare the AhR-associated proteins in transformed human fetal kidney cell lines in which AhR is constitutively active to those present in normal cells. Initial MALDI-TOF MS data analysis for samples derived from the GST-AhR transfected 293T cell line showed a number of protein bands (120 kDa, 90 kDa, 42 kDa and 35 kDa) and data base searches identified some of the proteins (HSP, BMALI), which are known to be associated with the AhR complex. We are in the process of sequencing additional peptides by MS/MS.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 环境中普遍存在的多环芳烃(PAH)和相关的二恶英的大部分活性都是由芳烃受体(AhR)介导的。AhR是Per/Arnt/Sim(PAS)转录因子家族的成员，被认为与神经发育、昼夜节律和血管生成有关。AhR在其N端的bHLH域含有一个DNA结合和蛋白质二聚化位点，在PAS同源结构域含有一个配体结合位点，核定位和输出序列，以及一个转录活性丰富的Q区。在与配体结合时，受体移位到细胞核中，并与至少一个辅助因子ArnT相关联。然后，激活的AhR复合体在特定的DNA结合部位与许多目标基因的启动子区域结合，导致基因转录。如果AhR在肿瘤细胞中具有结构性活性，并且其转录活性影响对这些细胞至关重要的功能，那么可以预期，在肿瘤细胞中，活性AhR复合体的组成将不同于未转化的细胞。这项研究的目的是鉴定和比较转化的人胎肾细胞系中的AhR相关蛋白，在这些细胞系中AhR具有结构性活性，而在正常细胞中存在这些蛋白。对转GST-AhR的293T细胞系样品进行的初步MALDI-TOF MS数据分析显示，有多条蛋白质带(120 kDa、90 kDa、42 kDa和35 kDa)，数据库搜索发现了一些已知与AhR复合体相关的蛋白质(HSP、BMALI)。我们正在用MS/MS对更多的多肽进行测序。