SV40 large T antigen up-regulates the retrotransposition frequency of viral-like 30 elements
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Noutsopoulos, D.
Vartholomatos, G.
Kolaitis, N.
Tzavaras, T.
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peer-reviewed
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J Mol Biol
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The regulation of non-autonomous retrotransposition is not known. A recombinant bearing a hygromycin gene and a viral-like 30 (VL30) retrotransposon tagged with an enhanced green fluorescent protein (EGFP) gene-based retrotransposition cassette was constructed and used for detection of retrotransposition events. Transfection of this recombinant produced retrotransposition events, detected both by EGFP fluorescence and PCR analysis, in hygromycin-selected clones of two established simian virus 40 (SV40)-transformed mouse NIH3T3 cell lines but not in normal NIH3T3 cells. The retrotransposition potential of this recombinant, as a provirus, was studied in stably transfected NIH3T3 clones. Transfection of these clones with either a wild-type or a mutant LE1135T SV40 large T antigen gene, not expressing small t protein, induced retrotransposition events at high frequencies as measured by fluorescence-activated cell scanning (FACS). In addition, measuring retrotransposition frequencies over a period of nine days following infection with isolated SV40 particles, revealed that the frequency of retrotransposition was time-dependent and induced as early as 24 h, increasing exponentially to high levels (>10(-2) events per cell per generation) up to nine days post-infection. Furthermore, ectopic expression of a cloned MoMLV-reverse transcriptase gene also produced retrotransposition events and suggested that the large T antigen most likely acted through induction of expression of endogenous reverse transcriptase genes. Our results show a direct correlation between SV40-cell transformation and VL30 retrotransposition and provide for the first time strong evidence that SV40 large T antigen up-regulates the retrotransposition of VL30 elements.
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Animals, Antigens, Polyomavirus Transforming/genetics/*physiology, Antigens, Viral, Tumor/genetics/physiology, Cell Line, Transformed, *Cell Transformation, Viral, Green Fluorescent Proteins/genetics/metabolism, Hygromycin B/metabolism, Mice, Moloney murine leukemia virus/enzymology, Mutation, NIH 3T3 Cells, Plasmids, RNA-Directed DNA Polymerase/genetics, Retroelements/genetics/*physiology, *Up-Regulation
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http://www.ncbi.nlm.nih.gov/pubmed/16859708
http://ac.els-cdn.com/S0022283606007637/1-s2.0-S0022283606007637-main.pdf?_tid=4d9be4aed7abe3be0d0dcd8bfec61d1e&acdnat=1333007296_7d09388614ee81602d0f8636f005ab6b
http://ac.els-cdn.com/S0022283606007637/1-s2.0-S0022283606007637-main.pdf?_tid=4d9be4aed7abe3be0d0dcd8bfec61d1e&acdnat=1333007296_7d09388614ee81602d0f8636f005ab6b
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en
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Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας. Τμήμα Ιατρικής