ROLE OF ETS1 IN LYMPHOID CELLS

ETS基因家族编码转录因子，与嘌呤结合- 丰富的基序(A/CGGAAGT/C)，并调节其启动子/增强子活性 细胞和病毒基因。ETS家族的成员包括：ETS1、ETS2、 ERG、ELK、ERGB/FLI、PU.1/SPII、ELF1、GABPalpha、PEA3、ER81和ER71 基因。为了了解ETS家族基因的功能 造血分化，特征良好的细胞系代表 红系、髓系和淋巴系已被用于研究ETS 通过RNA印迹分析进行表达。PEA3和ERG的表达分别为 在髓系前体细胞谱系中高于功能成熟的 髓系细胞。GABPalpha、ETS2 mRNAs在大多数细胞中表达 所研究的细胞系。然而，ETS1在淋巴组织中的表达要高得多 细胞比髓系或红系来源的细胞多。ERGB，GABPalpha， GABPβ和ETS2在胸腺细胞和脾B、T细胞中也有表达。 细胞。在有丝分裂原激活T细胞的过程中，ETS1和ERGB 表达减少。丹参对早幼粒细胞系HL60的刺激作用 佛波酯、GABPalpha和βmRNAs在早期阶段被诱导 差异化。因此，似乎ETS基因产物可能发挥了作用。 在细胞生长、分化和功能成熟中的作用 造血细胞。为了阐明ETS家族基因在人类免疫缺陷中的作用 T细胞，含DNA结合区的真核表达载体 并将不同形式的ETS1通过电转化法导入Jurkat T细胞。 在G418中筛选出了独立克隆。表征 这些克隆用于外源ETS基因产物的表达，及其 对靶基因的影响正在进行中。特别是流式细胞仪和RNA印迹 反映表面抗原表达模式和生长情况的分析 转基因细胞的特性(生长速度、成瘤性)、 淋巴因子(RNA和EL ISA)的产生和调节 ETS家族靶向的特定候选基因的活性 (RNA印迹、CAT分析、差异显示)，正在进行。

The ets family of genes encodes transcription factors, binds to purine- rich motifs (A/CGGAAGT/C), and regulates promoter/enhancer activities of cellular and viral genes. Members of the ets family include: ETS1, ETS2, ERG, ELK, ERGB/FLI, PU.1/SpiI, ELF1, GABPalpha, PEA3, ER81, and ER71 genes. To understand the function of the ets family of genes during hematopoietic differentiation, well-characterized cell lines representing erythroid, myeloid, and lymphoid lineages have been used to study ets expression by RNA blot analyses. The expression of PEA3 and ERG was higher in the myeloid precursor cell lineage than in functionally-mature myeloid cells. The GABPalpha, ETS2 mRNAs are expressed in most of the cell lines studied. However, ETS1 expression was much higher in lymphoid cells than in cells of myeloid or erythroid origin. ERGB, GABPalpha, GABPbeta and ETS2 was also expressed in thymocytes and splenic B- and T- cells. During T-cell activation by mitogens, both ETS1 and ERGB expression was reduced. Stimulation of promyelocytic cell line HL60 by phorbol esters, GABPalpha and beta mRNAs is induced during an early phase of differentiation. Therefore, it appears that ets gene products may play a role in cell growth, differentiation, and functional maturation of hematopoietic cells. To elucidate the role of the ets family of genes in T-cells, eukaryotic expression vectors containing the DNA-binding domain and variant forms of ETS1 were transfected into Jurkat T-cells by electro- poration, and independent clones were selected in G418. Characterization of these clones for expression of exogenous ets gene products, and their effect on target genes, is in progress. In particular, FACS and RNA blot analysis reflecting the expression pattern of surface antigen and growth properties of the transfected cells (growth rate, tumorigenesis), the production of lymphokines (RNA and ELISA), and the modulation of activities of specific candidate genes targeted by the ets family of genes (RNA blots, CAT assays, differential display), are being performed.