Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
We have developed a procedure for preparing extracts from nuclei of human tissue culture cells that directs accurate transcription initiation in vitro from class II promoters. Conditions of extraction and assay have been optimized for maximum activity using the major late promoter of adenovirus 2. The extract also directs accurate transcription initiation from other adenovirus promoters and cellular promoters. The extract also directs accurate transcription initiation from class III promoters (tRNA and Ad 2 VA).
D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000201:enzymology*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D009876:Operon; D011480:Protease Inhibitors / Q000494:pharmacology; D012319:RNA Polymerase II / Q000378:metabolism*; D012334:RNA, Neoplasm / Q000302:isolation & purification; D012343:RNA, Transfer / Q000235:genetics; D013696:Temperature; D013950:Thymus Gland / Q000201:enzymology; D014158:Transcription, Genetic / Q000187:drug effects
D011480:Protease Inhibitors; D012334:RNA, Neoplasm; D012343:RNA, Transfer; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases; D008274:Magnesium