T-antigen is the only detectable protein on the nucleosome-free origin region of isolated simian virus 40 minichromosomes.

A nucleosome-free region or nucleosome gap, containing the origin of replication and the transcriptional promoter elements, is observed on 20%-25% of the SV40 minichromosomes isolated at physiological ionic strength at late time during the infectious cycle. We found that this subpopulation of gapped minichromosomes was more sensitive to digestion with a variety of single-cut restriction enzymes than the rest of the minichromosomes. This increased digestibility of gapped minichromosomes allowed us to excise the gap region by concomitant digestion with Bgl I and Msp I. T-antigen was the only detectable protein bound to this isolated chromatin fragment. In particular no histones could be detected. The presence of T-antigen on the gap region was confirmed by immunoelectron microscopy. Most of the T-antigen appeared to be located on the late side of the Bgl I restriction enzyme site.