LoSardo J E, Cupelli L A, Short M K, Berman J W, Lenz J
Department of Genetics, Albert Einstein College of Medicine, Bronx, New York 10461.
J Virol. 1989 Mar;63(3):1087-94. doi: 10.1128/JVI.63.3.1087-1094.1989.
Transcriptional activities of the long terminal repeats (LTRs) of various murine leukemia viruses were tested in the cytotoxic T-cell lines CTLL-1 and CTLL-2. In contrast to T-lymphoma cells, in which the LTRs of T-lymphomagenic virus SL3-3 and Moloney murine leukemia virus are more active than those of other viruses, transcriptional activity in these mature, interleukin-2-dependent cells is not correlated with the specificity of viral leukemogenicity. Several approaches were used to investigate the molecular basis for LTR activity differences in lymphoma cells and mature cytotoxic T cells. Deletion analysis of the Moloney virus LTR showed that the direct repeats associated with enhancer activity have, at most, a slight effect on expression in CTLL-1 cells, whereas they stimulate expression six- to eightfold in T-lymphoma cells. This suggests that the mature T-cell line lacks one or more factors present in T-lymphoma cells that function to augment transcription from the Moloney murine leukemia virus LTR. We also used recombinant viral LTRs to investigate the role of the enhancer core element of SL3-3 in CTLL-1 and CTLL-2 cells. A one-base-pair difference between the core sequences of SL3-3 and nonleukemogenic Akv virus, which is important for SL3-3 activity in T-lymphoma cells, had no effect in these cells. The inability to distinguish the single-base-pair difference in expression assays was correlated with the absence of binding of a cellular factor, S-CBF, to the SL3-3 enhancer core in extracts of CTLL-1 and CTLL-2 nuclei. These studies may have implications for identification of the target cells for viral leukemogenesis, as well as for tracing of changes in the transcriptional machinery during T-lymphocyte differentiation.
在细胞毒性T细胞系CTLL-1和CTLL-2中检测了各种鼠白血病病毒长末端重复序列(LTR)的转录活性。与T淋巴瘤细胞不同,在T淋巴瘤细胞中,致T淋巴瘤病毒SL3-3和莫洛尼鼠白血病病毒的LTR比其他病毒的LTR更具活性,而在这些成熟的、依赖白细胞介素-2的细胞中,转录活性与病毒致白血病特异性无关。采用了几种方法来研究淋巴瘤细胞和成熟细胞毒性T细胞中LTR活性差异的分子基础。莫洛尼病毒LTR的缺失分析表明,与增强子活性相关的直接重复序列对CTLL-1细胞中的表达最多只有轻微影响,而它们在T淋巴瘤细胞中可刺激表达6至8倍。这表明成熟T细胞系缺乏T淋巴瘤细胞中存在的一种或多种增强莫洛尼鼠白血病病毒LTR转录的因子。我们还使用重组病毒LTR来研究SL3-3增强子核心元件在CTLL-1和CTLL-2细胞中的作用。SL3-3与非致白血病的Akv病毒核心序列之间的一个碱基对差异,对SL3-3在T淋巴瘤细胞中的活性很重要,但在这些细胞中没有影响。在表达分析中无法区分单碱基对差异与CTLL-1和CTLL-2细胞核提取物中细胞因子S-CBF与SL3-3增强子核心的结合缺失有关。这些研究可能对确定病毒致白血病的靶细胞以及追踪T淋巴细胞分化过程中转录机制的变化有启示。