Inhibition of the rous sarcoma virus long terminal repeat-driven transcription by in vitro methylation: different sensitivity in permissive chicken cells versus mammalian cells

Virology
J HejnarJ Svoboda

Abstract

Rous sarcoma virus (RSV) enhancer sequences in the long terminal repeat (LTR) have previously been shown to be sensitive to CpG methylation. We report further that the high density methylation of the RSV LTR-driven chloramphenicol acetyltransferase reporter is needed for full transcriptional inhibition in chicken embryo fibroblasts and for suppression of tumorigenicity of the RSV proviral DNA in chickens. In nonpermissive mammalian cells, however, the low density methylation is sufficient for full inhibition. The time course of inhibition differs strikingly in avian and mammalian cells: although immediately inhibited in mammalian cells, the methylated RSV LTR-driven reporter is fully inhibited with a significant delay after transfection in avian cells. Moreover, transcriptional inhibition can be overridden by transfection with a high dose of the methylated reporter plasmid in chicken cells but not in hamster cells. The LTR, v-src, LTR proviral DNA is easily capable of inducing sarcomas in chickens but not in hamsters. In contrast, Moloney murine leukemia virus LTR-driven v-src induces sarcomas in hamsters with high incidence. Therefore, the repression of integrated RSV proviruses in rodent cells is directed against the LTR.

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Citations

Aug 22, 2009·PLoS Pathogens·Jana BlazkovaIvan Hirsch
Feb 24, 2001·Proceedings of the National Academy of Sciences of the United States of America·J HejnarJ Svoboda
Jul 31, 2010·Methods : a Companion to Methods in Enzymology·Leonard ChávezEric Verdin
Nov 19, 2003·Biochemical and Biophysical Research Communications·Jirí HejnarJan Svoboda
Aug 16, 2005·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Chang Won ParkClifford J Steer
Jan 24, 2006·Experimental Cell Research·Magda MatouskováJirí Hejnar
Aug 22, 2007·Reproduction : the Official Journal of the Society for the Study of Fertility·Jirí KalinaPavel Trefil
Aug 30, 2013·Journal of Virology·Magda MatouskováJirí Hejnar
Sep 8, 2019·Viruses·Jiří Hejnar, Tomáš Ruml

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