Factors influencing the expression of endogenous reverse transcriptases and viral-like 30 elements in mouse NIH3T3 cells.
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.
Tzavaras
Theodore
T
University of Ioannina, Medical School, Laboratory of General Biology, GR-45 110 Ioannina, Greece. thtzavar@cc.uoi.gr
Eftaxia
Sofia
S
Tavoulari
Sotiria
S
Hatzi
Paraskevi
P
Angelidis
Charalambos
C
D000818:Animals; D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology; D015152:Blotting, Northern; D004247:DNA / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D003720:Densitometry; D003907:Dexamethasone / Q000494:pharmacology; D004054:Diethylstilbestrol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000494:pharmacology; D005799:Genes, Dominant; D051379:Mice; D041681:NIH 3T3 Cells; D010957:Plasmids / Q000378:metabolism; D011374:Progesterone / Q000494:pharmacology; D012313:RNA / Q000737:chemistry; D012194:RNA-Directed DNA Polymerase / Q000096:biosynthesis* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D018931:Antineoplastic Agents, Hormonal; D017931:DNA Primers; D016159:Tumor Suppressor Protein p53; D011374:Progesterone; D004958:Estradiol; D012313:RNA; D004054:Diethylstilbestrol; D003907:Dexamethasone; D004247:DNA; D012194:RNA-Directed DNA Polymerase