The SV40 nucleosome-free region is detected throughout the virus life cycle.
The structures of SV40 intracellular chromatin complexes and of extracellular virus particles were examined by photolabeling with a radioactive psoralen derivative in order to determine the fate of the exposed origin region during the virus life cycle. We have previously shown that the origin region of intracellular SV40 chromatin is preferentially accessible to psoralen derivatives in vivo, whereas psoralen adducts are uniformly distributed when purified virus particles are photoreacted. We demonstrate here that when virion is photoreacted prior to a freeze-thaw cycle, the exposed regulatory region detected in intracellular nucleoprotein complexes is also found in mature virus particles. In contrast, if the virion is frozen and thawed prior to the photoreaction, the origin is not preferentially exposed to photoaddition. Virus particles that have not been subjected to a freeze-thaw cycle were found to exhibit preferential labeling in the origin region whether they were irradiated intracellularly, in culture medium, or following purification. Banding the virus in CsCl had no significant effect on the relative accessibility of the origin region to psorealen. Our findings indicate that the open regulatory region found on intracellular SV40 chromatin persists throughout the virus life cycle.
Kondoleon
S K
SK
Department of Microbiology and Immunology, Oregon Health Sciences University, Portland 97201.
Kurkinen
N A
NA
Hallick
L M
LM
D000818:Animals; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002843:Chromatin / Q000648:ultrastructure*; D004278:DNA, Superhelical / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D005615:Freezing; D011564:Furocoumarins; D009698:Nucleoproteins / Q000235:genetics; D009707:Nucleosomes / Q000648:ultrastructure*; D015183:Restriction Mapping; D013539:Simian virus 40 / Q000235:genetics / Q000254:growth & development* / Q000648:ultrastructure; D014771:Virion / Q000235:genetics / Q000254:growth & development / Q000648:ultrastructure
D002843:Chromatin; D004278:DNA, Superhelical; D004279:DNA, Viral; D011564:Furocoumarins; D009698:Nucleoproteins; D009707:Nucleosomes