Effects of DNA methylation on specific transcription by RNA polymerase II in vitro.
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 degrees.
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Molloy
D000818:Animals; D002474:Cell-Free System; D002645:Chickens; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D004262:DNA Restriction Enzymes; D005796:Genes; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007633:Keratins / Q000235:genetics*; D007700:Kinetics; D008745:Methylation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism*; D013698:Templates, Genetic; D014158:Transcription, Genetic
D007633:Keratins; D004247:DNA; D012319:RNA Polymerase II; D004262:DNA Restriction Enzymes