SV40 virus particles lack a psoralen-accessible origin and contain an altered nucleoprotein structure.
The nucleoprotein structure of SV40 virions was examined by photolabeling purified virus with the radioactive psoralen derivative hydroxymethyltrimethylpsoralen (HMT). Unlike SV40 chromatin in situ, the viral origin region is not preferentially accessible to drug addition. The ratio of the distribution of radioactivity in the DNA restriction fragments of virion DNA to that of purified SV40 DNA demonstrates that the photoadducts are positioned similarly on the circular molecule in both samples. Virion purified from infected cells was also analyzed for the presence of an open region and found to exhibit the same pattern of [3H]HMT addition as mature extracellular virion. The nucleosome-free region detected at the SV40 replication origin in intracellular minichromosomes is not present in either population of intact virus particles. We also examined the level of drug addition obtained when purified virion or SV40-infected cells were treated with saturating doses of [3H]HMT. Marked differences in the plateau levels of bound drug indicate that an altered nucleoprotein structure exists in SV40 virions that does not protect the DNA from photoaddition to the same extent as do the nucleosomes of intracellular SV40 DNA.
D000818:Animals; D002460:Cell Line; D055598:Chemical Phenomena; D002621:Chemistry; D002522:Chlorocebus aethiops; D002843:Chromatin / Q000032:analysis*; D004261:DNA Replication; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000032:analysis* / Q000378:metabolism; D013539:Simian virus 40 / Q000032:analysis*; D014307:Trioxsalen / Q000031:analogs & derivatives / Q000378:metabolism; D014771:Virion / Q000032:analysis*; D014779:Virus Replication
D002843:Chromatin; D004279:DNA, Viral; C016507:hydroxymethyltrioxsalen; D004262:DNA Restriction Enzymes; D014307:Trioxsalen