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Effects of DNA methylation on specific transcription by RNA polymerase II in vitro

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Abstract

In vitro transcription in a HeLa cell lysate by RNA polymerase II directed by a chicken feather keratin gene promotor has been studied using unmethylated template DNA and DNA methylated in vitro by HpaII methylase. The efficiency of specific gene transcription from methylated DNA was dependent on topology of the input DNA, the most significant effect being complete inhibition of transcription from one template which contained three methylation sites, one just 5′ and two greater than 500 bases 3′ to the site of transcription initiation. The inhibition of transcription depends on a factor(s) which is variably present in lysate preparations and is labile on storage at -70°.

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  1. CSIRO Division of Molecular Biology, P.O. Box 184, 2113, North Ryde, N.S.W., Australia

    P. L. Molloy

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  1. P. L. Molloy
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Molloy, P.L. Effects of DNA methylation on specific transcription by RNA polymerase II in vitro . Mol Biol Rep 11, 13–17 (1986). https://doi.org/10.1007/BF00417589

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  • DOI: https://doi.org/10.1007/BF00417589

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