Tzavaras Theodore, Eftaxia Sofia, Tavoulari Sotiria, Hatzi Paraskevi, Angelidis Charalambos
University of Ioannina, Medical School, Laboratory of General Biology, GR-45 110 Ioannina, Greece.
Int J Oncol. 2003 Oct;23(4):1237-43.
Retroviral reverse transcriptase (RT) plays a definite role in retroviral life cycle and is essential for the process of retrotransposition. We investigated the RNA expression of endogenous reverse transcriptases (enRTs) in the NIH3T3 mouse genome using, as a probe, a mixture of RT-PCR generated reverse transcriptase products potentially detecting a large number of RTs following treatment with different agents. We found that the expression of enRTs is induced approximately 500-fold following 5'-azacytidine-treatment. Amongst steroid hormones used such as estradiol, diethylstilbestrol, progesterone and dexamethasone only the latter was effective in inducing enRTs up to 4-fold at a concentration of 10(-7) M. Expression of a mouse dominant-negative form of p53 protein in cell clones resulted in induction of 20- to 50-fold, whereas C2-ceramide in a 4-fold induction at concentrations of 20-80 micro M. In a parallel analysis, the respective expression of the transposable viral-like 30 elements (VL30s) was also measured. Their expression was induced up to 50-fold by 5'-azacytidine, overexpression of the p53 gene and C2-ceramide at 80 micro M. It was also induced approximately 3- to 5-fold following estradiol, diethylstilbestrol or progesterone treatment and 30-fold by dexamethasone. Collectively, our results suggest that such stimuli inducing enRTs might play a role in the activation of transcription and retrotransposition of VL30.
逆转录病毒逆转录酶(RT)在逆转录病毒生命周期中发挥着明确作用,对于逆转座过程至关重要。我们使用RT-PCR产生的逆转录酶产物混合物作为探针,研究了NIH3T3小鼠基因组中内源性逆转录酶(enRTs)的RNA表达,该混合物在不同试剂处理后可能检测到大量逆转录酶。我们发现,5'-氮杂胞苷处理后enRTs的表达诱导了约500倍。在所使用的类固醇激素中,如雌二醇、己烯雌酚、孕酮和地塞米松,只有后者在浓度为10^(-7) M时能有效诱导enRTs高达4倍。在细胞克隆中表达小鼠显性阴性形式的p53蛋白导致诱导20至50倍,而20至80微摩尔浓度的C2-神经酰胺诱导4倍。在平行分析中,还测量了转座病毒样30元件(VL30s)的各自表达。它们的表达在5'-氮杂胞苷、p53基因过表达和80微摩尔的C2-神经酰胺作用下诱导高达50倍。在雌二醇、己烯雌酚或孕酮处理后也诱导约3至5倍,地塞米松诱导30倍。总体而言,我们的结果表明,这种诱导enRTs的刺激可能在VL30的转录激活和逆转座中发挥作用。