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.