title: FACT, a factor that facilitates transcript elongation through nucleosomes.
abstract: The requirements for transcriptional activation by RNA polymerase II were examined using chromatin templates assembled in vitro and a transcription system composed of the human general transcription factors and RNA polymerase II. Activator-induced, energy-dependent chromatin remodeling promoted efficient preinitiation complex formation and transcription initiation, but was not sufficient for productive transcription. Polymerases that initiated transcription on remodeled chromatin templates encountered a block to transcription proximal to the promoter. Entry into productive transcription required an accessory factor present in HeLa cell nuclear extract, FACT (facilitates chromatin transcription), which we have purified. FACT acts subsequent to transcription initiation to release RNA polymerase II from a nucleosome-induced block to productive transcription. The biochemical properties and polypeptide composition of FACT suggest that it is a novel protein factor that facilitates transcript elongation through nucleosomes.
authors.affiliation: Howard Hughes Medical Institute, Department of Biochemistry, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway 08854, USA.
authors.forename: G
authors.initials: G
authors.lastname: Orphanides
authors.forename: G
authors.initials: G
authors.lastname: LeRoy
authors.forename: C H
authors.initials: CH
authors.lastname: Chang
authors.forename: D S
authors.initials: DS
authors.lastname: Luse
authors.forename: D
authors.initials: D
authors.lastname: Reinberg
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D001665:Binding Sites / Q000502:physiology; D002474:Cell-Free System / Q000737:chemistry / Q000378:metabolism; D002843:Chromatin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins; D006367:HeLa Cells; D006609:High Mobility Group Proteins / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D008875:Middle Aged; D009687:Nuclear Proteins / Q000378:metabolism; D056489:Nucleoplasmins; D009707:Nucleosomes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic / Q000235:genetics / Q000502:physiology*; D035602:Transcriptional Elongation Factors
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D009687:Nuclear Proteins; D056489:Nucleoplasmins; D009707:Nucleosomes; D010750:Phosphoproteins; C073537:SSRP1 protein, human; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; D012313:RNA; D000255:Adenosine Triphosphate; D012319:RNA Polymerase II