Black A C, Luo J, Chun S, Tabibzadeh S
Department of Medicine, UCLA School of Medicine 90095-1678, USA.
Virus Genes. 1997;15(2):105-17. doi: 10.1023/a:1007906823269.
Human T-cell leukemia virus type I (HTLV-I) causes adult T-cell leukemia/lymphoma (ATLL). HTLV Tax, the viral transcriptional activator, can activate a variety of cellular genes. HTLV-mediated T-cell transformation, however, may involve additional viral proteins expressed from singly- as well as doubly-spliced viral mRNA. To determine the combined effect of these viral proteins on cellular gene expression in Jurkat T-cells, we derived stable transfectants that constitutively express the HTLV-I pX and env regions (J3.9). J3.9 cells show substantially increased mRNA levels of egr-1 and c-jun but no induction of either CD25 or GM-CSF by Northern blotting. This pattern corresponded to the activation of an egr-1 but not a GM-CSF promoter-driven reporter construct in transient gene expression assays. In DNA electrophoretic mobility shift assays (EMSA), nuclear extract from J3.9 cells has significantly increased binding to CRE and SRE but not nuclear factor kappa B (NF kappa B) DNA oligos, as compared to J-Neo cell extract. These results suggest that low level expression of pX and env region gene products in Jurkat T-cells stimulates persistent activation of CRE- and SRE- but not NF kappa B-induced cellular genes.
人类I型T细胞白血病病毒(HTLV-I)可引发成人T细胞白血病/淋巴瘤(ATLL)。病毒转录激活因子HTLV Tax能够激活多种细胞基因。然而,HTLV介导的T细胞转化可能涉及从单剪接和双剪接病毒mRNA表达的其他病毒蛋白。为了确定这些病毒蛋白对Jurkat T细胞中细胞基因表达的联合作用,我们构建了稳定转染细胞系,其组成性表达HTLV-I的pX和env区域(J3.9)。通过Northern印迹法检测发现,J3.9细胞中egr-1和c-jun的mRNA水平显著升高,但CD25或GM-CSF均未被诱导。这种模式与瞬时基因表达试验中egr-1启动子驱动的报告基因构建体的激活相对应,但GM-CSF启动子驱动的报告基因构建体未被激活。在DNA电泳迁移率变动分析(EMSA)中,与J-Neo细胞提取物相比,J3.9细胞的核提取物与CRE和SRE的结合显著增加,但与核因子κB(NFκB)DNA寡核苷酸的结合没有增加。这些结果表明,Jurkat T细胞中pX和env区域基因产物的低水平表达刺激了CRE和SRE诱导的细胞基因的持续激活,但未刺激NFκB诱导的细胞基因的激活。