Transcription of fractionated mammalian chromatin by mammalian ribonucleic acid polymerase. Demonstration of temperature-dependent rifampicin-resistant initiation sites in euchromatin deoxyribonucleic acid.
The chromatin fractionation method of Frenster et al. (1963) as modified by Leake et al. (1972) was used to prepare fragments of euchromatin from rat liver nuclei. These remain soluble in 5mm-MgCl(2), and contain DNA of maximum mol.wt. 1x10(6)-2x10(6). The fragments were separated from condensable chromatin on a sucrose gradient. Euchromatin contains endogenous DNA-dependent RNA polymerase, and most of the nascent RNA labelled in vivo or in vitro. Euchromatin fragments allow initiation of transcription by added purified rat liver form-B RNA polymerase and contain temperature-dependent rifampicin-resistant initiation sites for the form-B enzyme. These findings indicate that transcription of the euchromatin regions of interphase chromosomes is not initiated in condensed chromatin, but is initiated within the euchromatin stretches. Condensable chromatin also contains most of these activities, but is not associated with nascent RNA.
D000818:Animals; D002467:Cell Nucleus / Q000032:analysis; D005591:Chemical Fractionation; D002843:Chromatin / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D004247:DNA / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000032:analysis / Q000378:metabolism*; D004351:Drug Resistance; D066298:In Vitro Techniques; D008099:Liver / Q000032:analysis; D008970:Molecular Weight; D011506:Proteins / Q000032:analysis; D051381:Rats; D012293:Rifampin / Q000378:metabolism; D013010:Sonication; D013696:Temperature; D014158:Transcription, Genetic; D014316:Tritium; D014529:Uridine
D002843:Chromatin; D011506:Proteins; D014316:Tritium; D004247:DNA; D012321:DNA-Directed RNA Polymerases; D012293:Rifampin; D014529:Uridine