Intracellular forms of simian virus 40 nucleoprotein complexes. V. Enrichment for "active" simian virus 40 chromatin by differential precipitation with Mg2+.
A fraction (approximately 20%) of 70S simian virus (SV40) chromatin isolated from a sucrose gradient-purified extract of SV40-infected CV-1 cells was found to be precipitated by incubation with a buffer containing 10 mM Mg2+ at 37 degrees C. The fraction containing the Mg2+-insoluble SV40 chromatin was found also to be enriched for the following: (i) DNA replication intermediates as analyzed by short pulse with [3H]thymidine; (ii) transcription complexes as assayed by in vitro synthesis of SV40-specific RNA; (iii) newly synthesized T antigen; (iv) phosphorylated T antigen as analyzed by both in vitro and in vivo phosphorylation studies; and (v) ATP-binding activity of T antigen as assayed by affinity labeling with oxidized [alpha-32P]ATP.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000956:Antigens, Viral / Q000302:isolation & purification; D000957:Antigens, Viral, Tumor; D002460:Cell Line; D002843:Chromatin / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D004261:DNA Replication; D005590:Fractional Precipitation; D000882:Haplorhini; D008274:Magnesium; D010766:Phosphorylation; D013539:Simian virus 40 / Q000032:analysis* / Q000276:immunology; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000302:isolation & purification*
D000956:Antigens, Viral; D000957:Antigens, Viral, Tumor; D002843:Chromatin; D014764:Viral Proteins; D000255:Adenosine Triphosphate; D008274:Magnesium