C-ETS GENE EXPRESSION DURING CELL PROLIFERATION AND DIFFERENTIATION

细胞增殖和分化过程中的 C-ETS 基因表达

• 批准号: 3874673
• 负责人: T S PAPAS
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3874673
• 关键词: CD4 molecule; CD8 molecule; T cell receptor; T lymphocyte; antibody specificity; antiserum; cell differentiation; cell growth regulation; complementary DNA; crosslink; developmental genetics; fibroblasts; gene expression; genetic markers; helper T lymphocyte; human tissue; ionomycin; leukocyte activation /transformation; lymphokines; messenger RNA; mitogens; molecular cloning; monoclonal antibody; neoplasm /cancer genetics; neoplasm /cancer immunodiagnosis; neoplastic cell culture for noncancer research; nucleoproteins; oligopeptides; oncogenes; oncoproteins; phytohemagglutinins; posttranslational modifications; protein kinase C; protooncogene; radioimmunoassay; second messengers; tumor antigens; western blottings

The ETS1 and ETS2 gene expression has been studied in purified human T-cells activated by either cross-linking of the T-cell receptor-CD3 complex on their cell surface or by direct stimulation with phorbol esters and ionomycin. The quiescent T-cells express high levels of ETS1 and undetectable levels of ETS2 mRNA and proteins. Upon T-cell activation, ETS2 gene products are induced while ETS1 mRNA and proteins decrease to very low levels. Late after stimulation, ETS1 mRNA is reinduced and maintained at a high level, while ETS2 gene expression decreases to very low levels. These results suggest that in human T-cells ETS2 gene products are associated with cellular activation and proliferation, while ETS1 gene products are preferentially expressed in a quiescent state. The reciprocal regulation of ETS1 and ETS2 genes is observed only in T-cells, but not in B- and non-lymphoid cells. Experiments are in progress to deprive ETS1 and ETS2 proteins to understand the specific role of c-ets gene products in thymocyte and T-cell differentiation, maturation, activation and proliferation. The pan-ets monoclonal antibody detects c-ets-1, c-ets-2, c-erg proteins in radioimmunoprecipitation assays, while c-ets-1 and additional 55 kDa proteins are detected by Western blot analysis. Multiple murine ets-1 (58,54,50 and 40 kDa) and ets-2 (56 kDa) have been identified as ets-1 and ets-2 gene-encoded products. Only 58, 54 kDa ets-1 and ets-2 nuclear proteins are hyperphosphorylated by Ca2+ mediated events. The origin of multiple ets-1 proteins, their post-translational modification and their functional significance are under investigation. The ERG genes are expressed at very low levels and their expression is restricted to a few cell types. They are expressed at high levels in immature rather than in mature myeloid cells. Different regions of ERG cDNA have been expressed in E. coli and purified proteins are being used to generate both polyclonal and monoclonal antibodies.

ETS 1和ETS 2基因的表达已经在纯化的人 通过T细胞受体-CD 3的交联激活的T细胞 复合物在其细胞表面或通过直接刺激与佛波醇酯 和离子霉素。静止T细胞表达高水平的ETS 1， 无法检测到的ETS 2 mRNA和蛋白水平。T细胞激活后，ETS 2 基因产物被诱导，而ETS 1 mRNA和蛋白质下降到非常低 程度.刺激后晚期，ETS 1 mRNA被重新诱导并维持在一个恒定的水平。 高水平，而ETS 2基因表达降低到非常低的水平。这些 结果表明，在人类T细胞中，ETS 2基因产物与 与细胞活化和增殖，而ETS 1基因产物， 优先在静止状态下表达。的相互调节 ETS 1和ETS 2基因仅在T细胞中观察到，而在B-和 非淋巴细胞。实验正在进行中，以去除ETS 1和ETS 2 蛋白质，以了解c-ets基因产物在 胸腺细胞和T细胞分化、成熟、活化和 增殖pan-ets单克隆抗体检测c-ets-1，c-ets-2， c-erg蛋白在放射免疫沉淀分析中，而c-ets-1和 通过蛋白质印迹分析检测另外的55 kDa蛋白质。多 已经鉴定了鼠ets-1（58、54、50和40 kDa）和ets-2（56 kDa 作为ETS-1和ETS-2基因编码的产物。只有58，54 kDa ets-1和ets-2 核蛋白被Ca 2+介导的事件过度磷酸化。的 多种ets-1蛋白的起源、它们的翻译后修饰 其功能意义正在研究中。ERG基因 以非常低的水平表达，并且它们的表达仅限于 几种细胞类型。它们在未成熟的而不是成熟的细胞中高水平表达。 在成熟的骨髓细胞中。ERG cDNA的不同区域已经表达， 在大肠大肠杆菌和纯化的蛋白质被用来产生多克隆的 和单克隆抗体。