Linear association between cellular DNA and Epstein-Barr virus DNA in a human lymphoblastoid cell line.
High-molecular-weight DNA from cell line Raji (derived from Burkitt's lymphoma), which contains 50-60 copies of Epstein-Barr virus DNA per cell, was fractionated in neutral solution by several cycles of CsCl gradient centrifugation in fixed-angle rotors. Under the fractionation conditions used, intact Epstein-Barr virus DNA from virus particles can be separated from the less-dense cellular DNA. In contrast, a large proportion of the intrinsic Epstein-Barr virus DNA component of Raji cells remains associated with cellular DNA, as determined by nucleic acid hybridization. This interaction, which is resistant to Pronase and phenol treatment, is not the result of aggregation. When the molecular weight of Raji DNA is reduced by hydrodynamic shear, the amount of virus DNA associated with cell DNA decreases. However, some virus DNA still remains bound to fragments of cellular DNA after shearing. The association is completely destroyed in alkaline solution. Molecular weight analysis of Raji DNA after denaturation showed that the alkali-induced release of Epstein-Barr virus DNA was specific and not the result of random single-strand breaks. These data indicate that Epstein-Barr virus DNA is linearly integrated into Raji cell DNA by alkali-labile bonds.
D002051:Burkitt Lymphoma / Q000382:microbiology; D002250:Carbon Radioisotopes; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004269:DNA, Bacterial / Q000378:metabolism; D004273:DNA, Neoplasm / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D004279:DNA, Viral / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D004854:Herpesvirus 4, Human; D006801:Humans; D006868:Hydrolysis; D008970:Molecular Weight; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D010761:Phosphorus Radioisotopes; D011402:Pronase; D012367:RNA, Viral / Q000378:metabolism; D014316:Tritium
D002250:Carbon Radioisotopes; D004269:DNA, Bacterial; D004273:DNA, Neoplasm; D004279:DNA, Viral; D010761:Phosphorus Radioisotopes; D012367:RNA, Viral; D014316:Tritium; D011402:Pronase