Integration, loss, and reacquisition of defective viral DNA in SV40-transformed mouse cell lines.
We have examined the state of viral DNA in a set of SV40-transformed mouse cell lines. Using restriction enzymes which cut SV40 DNA in one place, we demonstrate that anchorage-independent SV40-transformed mouse cells commonly contain one or more detectable defective monomers of integrated viral DNA. The defective viral DNA in one of these cell lines, SV101, was extensively mapped using single and double enzyme digests. The results of this analysis indicate that SV101 contains nondefective viral DNA as well as defective viral DNA of the following sizes: 5.0, 4.3, 3.7, 3.4, and 1.5 kb. Three of these defective monomers (4.3, 3.7, and 1.5 kb) preserve the amino terminal exon of large T antigen, and two monomers (4.3, and 3.7 kb) preserve the little t coding region. Anchorage-dependent subclones of SV101 preferentially lose the defective viral DNA, while retaining an intact SV40 early region and the ability to express lytic-sized large and small T antigens. Despite a considerable amount of viral DNA rearrangement which accompanies subcloning, anchorage-independent subclones of SV101 retain defective viral DNA, especially the 4.3- and 3.7-kb monomers. Also, when an anchorage-independent subclone is selected from an anchorage-dependent revertant of SV101, it reacquires defective viral DNA, although of a size not seen in SV101. We conclude that defective viral DNA plays a role in generating the anchorage-independent phenotype. In earlier studies, we have reported that anchorage-transformed mouse lines contain a variant (100kDa) T antigen. The possible role of defective viral DNA in generating this T antigen is discussed.
D000818:Animals; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D004261:DNA Replication; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics*; D051379:Mice; D009154:Mutation; D009693:Nucleic Acid Hybridization; D013539:Simian virus 40 / Q000235:genetics*; D014779:Virus Replication
D004279:DNA, Viral; D004262:DNA Restriction Enzymes