Open AccessFormation of transcribing mononucleosome-eukaryotic RNA polymerase II complexesin vitroas a simple model of active chromatin
Author(s) -
Kei Sakuma,
Yoshihiro Matsumura,
Tatsuo Senshu
Publication year - 1984
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/12.3.1415
Subject(s) - biology , polymerase , rna polymerase ii , microbiology and biotechnology , linker dna , rna polymerase , chromatin , dna clamp , nucleosome , dna polymerase i , transcription (linguistics) , dna , rna polymerase i , rna polymerase iii , dna polymerase , transcription bubble , biochemistry , rna , rna dependent rna polymerase , reverse transcriptase , gene expression , promoter , gene , linguistics , philosophy
Mononucleosomes obtained from cultured mouse hepatoma cells were incubated with RNA polymerase II from wheat germ. No free DNA was liberated as available templates under the experimental condition employed. Size analysis of the transcripts showed that the polymerase initiated transcription from either terminus and read through the DNA template of mononucleosomes. Sucrose density gradient centrifugation of the reaction mixture resolved mononucleosome-polymerase complexes from free materials. The complexes were characterized by the enrichment of DNA fragments containing the nucleosome linker region, the presence of H1 histone, and the increased susceptibility to DNase I. Both the complexes formed in the presence and absence of precursor nucleotides were susceptible. These suggest that RNA polymerase II prefers to bind to the linker region, and the polymerase-bound nucleosomes are structurally altered. The data were discussed in context with possible mechanisms of transcription of the nucleosome structure.