Histone hyperacetylation during SV40 transcription is regulated by p300 and RNA polymerase II translocation.
The effects of the RNA polymerase II (RNAPII) translocation inhibitors alpha amanitin and 5,6-dichloro-1-beta-D-ribobenzimidazole (DRB) and an siRNA targeting p300 on the presence of RNAPII, p300, hyperacetylated H4 and H3 and unmodified H4 and H3 in transcribing simian virus 40 (SV40) minichromosomes were determined. Following treatment with alpha amanitin we observed a profound reduction in the occupancy of the promoter by RNAPII, the loss of p300 from chromatin fragments containing RNAPII, and an increase in the amount of unmodified H4 and H3 associated with the RNAPII. Treatment with DRB had little effect on the presence of RNAPII or p300 but also resulted in a significant increase in the amount of unmodified H4 and H3 present in chromatin fragments associated with RNAPII. Following treatment with a p300 small interfering RNA (siRNA), we observed a significant decrease in late transcription and a corresponding reduction in the amounts of p300 and hyperacetylated histones associated with the transcribing SV40 minichromosomes. We conclude that in transcribing SV40 minichromosomes histone hyperacetylation and deacetylation is dependent upon the presence of p300 and an as yet unknown histone deacetylase associated with the RNAPII complex that occurs coordinately as the RNAPII complex moves through a nucleosome.
Department of Biochemistry and Molecular Biology, University of North Dakota, Grand Forks, ND 58203, USA.
Lata
L
Balakrishnan
Barry
B
Milavetz
D000107:Acetylation; D000546:Amanitins / Q000235:genetics; D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002460:Cell Line; D002875:Chromosomes / Q000235:genetics; D015536:Down-Regulation; D015967:Gene Expression Regulation, Viral; D016679:Genome, Viral / Q000235:genetics; D000882:Haplorhini; D051548:Histone Acetyltransferases / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D021381:Protein Transport; D012319:RNA Polymerase II / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D012263:Ribonucleosides / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics*; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor
D000546:Amanitins; D018797:Cell Cycle Proteins; D051548:Histone Acetyltransferases; D006657:Histones; D012319:RNA Polymerase II; D034741:RNA, Small Interfering; D012263:Ribonucleosides; D014157:Transcription Factors; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor