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.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的首席研究员可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 环境中普遍存在的多环芳烃 (PAH) 和相关二恶英的大部分活性都是由 AhR（芳烃受体）介导的。 AhR 是转录因子 Per/ARNT/Sim (PAS) 家族的成员，已知与神经发育、昼夜节律周期和血管生成相关。 AhR 在其 N 端 bHLH 结构域中包含 DNA 结合和蛋白质二聚化位点、PAS 同源结构域中的配体结合位点、核定位和输出序列以及转录活性丰富的 Q 结构域。配体结合后，受体易位到细胞核中并与至少一种辅助因子 ARNT 结合。激活的 AhR 复合物然后在特定的 DNA 结合位点结合许多靶基因的启动子区域，导致基因转录。如果AhR在肿瘤细胞中具有组成型活性并且其转录活性影响对这些细胞至关重要的功能，则可以预期肿瘤细胞中的活性AhR复合物的组成将不同于其未转化的对应物。本研究的目的是鉴定和比较转化人胎儿肾细胞系中的 AhR 相关蛋白，其中 AhR 与正常细胞中存在的蛋白具有组成型活性。对源自 GST-AhR 转染的 293T 细胞系的样品进行的初始 MALDI-TOF MS 数据分析显示了许多蛋白质条带（120 kDa、90 kDa、42 kDa 和 35 kDa），并且数据库搜索鉴定了一些蛋白质（HSP、 BMALI)，已知其与 AhR 复合体相关。我们正在通过 MS/MS 对其他肽进行测序。