Unwinding of chromatin by the SV40 large T antigen DNA helicase.
We have analysed the unwinding of nucleosomally organized DNA by simian virus 40 large tumour (T) antigen. Isolated T antigen can bind to existing nucleosome cores containing the viral replication origin sequence, which results in displacement of the histone octamer and unwinding of the DNA. However, specific binding to nucleosome cores is salt sensitive and nearly completely blocked under ionic conditions that otherwise support DNA replication. Once started, the progressing T antigen helicase, like an elongating RNA polymerase, is not further repressed by histone octamers, irrespective of the presence or absence of linker histone H1. Disruption of the nucleosomal structure in the process of unwinding may be assisted by the demonstrated interaction of the hexameric T antigen complex with histone proteins H1 and H3. Finally, our studies reveal the inability of topoisomerase I and/or II to continually relieve the superhelical tension of covalently closed circular minichromosomes as generated during their unwinding by T antigen. This may indicate that chromatin relaxation during the process of DNA replication can only be efficiently performed by a topoisomerase that is (trans)activated by other factors.
Ramsperger
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U
Fakultät für Biologie, Universität Konstanz, Germany.
Stahl
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H
D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry*; D002843:Chromatin / Q000737:chemistry*; D002875:Chromosomes / Q000737:chemistry; D004247:DNA / Q000096:biosynthesis; D004265:DNA Helicases / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000737:chemistry; D005260:Female; D006657:Histones / Q000737:chemistry; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000737:chemistry; D009865:Oocytes; D018741:Replication Origin / Q000235:genetics; D012965:Sodium Chloride / Q000737:chemistry; D018411:Spodoptera; D014982:Xenopus laevis
D000952:Antigens, Polyomavirus Transforming; D002843:Chromatin; D004268:DNA-Binding Proteins; D006657:Histones; D009707:Nucleosomes; D012965:Sodium Chloride; D000255:Adenosine Triphosphate; D004247:DNA; D004265:DNA Helicases