Assembly of RNA polymerase II preinitiation complexes before assembly of nucleosomes allows efficient initiation of transcription on nucleosomal templates.
We have previously shown that assembly of nucleosomes on the DNA template blocks transcription initiation by RNA polymerase II in vitro. In the studies reported here, we demonstrate that assembly of a complete RNA polymerase II preinitiation complex before nucleosome assembly results in nucleosomal templates which support initiation in vitro as efficiently as naked DNA. Control experiments prove that our observations are not the result of slow displacement of nucleosomes by the transcription machinery during chromatin assembly, nor are they an artifact of inefficient nucleosome deposition on templates already bearing an RNA polymerase. Thus, the RNA polymerase II preinitiation complex appears to be resistant to disruption by subsequent nucleosome assembly.
Knezetic
J A
JA
Department of Biochemistry and Molecular Biology, University of Cincinnati College of Medicine, Ohio 45267-0522.
Jacob
G A
GA
Luse
D S
DS
D002843:Chromatin / Q000378:metabolism; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D009707:Nucleosomes / Q000378:metabolism*; D010957:Plasmids; D012313:RNA / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012319:RNA Polymerase II / Q000378:metabolism*; D013698:Templates, Genetic; D014158:Transcription, Genetic
D002843:Chromatin; D009707:Nucleosomes; D012313:RNA; D012319:RNA Polymerase II